Anti-inflammatory effects of quinolinyl analog of resveratrol targeting TLR4 in MCAO/R ischemic stroke rat model.

BACKGROUND: Among adults, stroke is the main causes of mortality and permanent disability. Neuroinflammation is one of the main causes of stoke-mediated neuronal death. Our previous study revealed that (E)-5-(2-(Quinolin-4-yl) vinyl) benzene-1, 3-diol (RV01), a quinolinyl analog of resveratrol, inhibits microglia-induced neuroinflammation and safeguards neurons from inflammatory harm. The preventive role of RV01 in ischemic stroke and its underlying cellular mechanisms and molecular targets remain poorly understood. PURPOSE: To investigate whether RV01 alleviates ischemia-reperfusion (I/R) injury by inhibiting microglia-mediated neuroinflammation and determine the potential molecular mechanisms and targets by which RV01 inhibits the I/R-mediated microglia activation. METHODS: Rat middle cerebral artery occlusion and reperfusion (MCAO/R) and BV-2 or primary microglial cells oxygen-glucose deprivation and reperfusion (OGD/R) models were established. The neurological behavior scores, 2, 3, 5-triphenyl tetrazolium chloride staining and immunofluorescence were used to detect the neuroprotective effect of RV01 in the MCAO/R rats. In addition, the mRNA expression levels of IL-6, TNF-α, and IL-1ß were detected to reveal the antineuroinflammatory effect of RV01. Moreover, a western blot assay was performed to explore the protein expression changes in NF-κB-mediated neuroinflammation. Finally, we identified TLR4 as an RV01 target through molecular docking, drug sensitivity target stability analysis, cellular thermal shift analysis, and surface plasmon resonance techniques. RESULTS: RV01 reduced the infarct volume and neurological deficits, increased the rotarod duration, and decreased the number of rightward deflections in the MCAO/R rats. RV01 inhibited the NF-κB signaling pathway in vitro and in vivo, as demonstrated by the reduction in the transcription factor p65-mediated expression of several inflammatory factors including IL-6, TNF-α, and IL-1ß. Further studies showed that its protective effect was associated with targeting the TLR4 protein. Notably, the anti-inflammatory effect of RV01 was markedly reinforced by the TLR4 knockdown, but inhibited by the overexpression of TLR4. Results revealed that the conditioned medium derived from the RV01-treated BV-2 cells significantly decreased the OGD/R-mediated neuronal damage. CONCLUSION: Our results are the first to reveal the protective effects of RV01 on cerebral ischemia, depending on its inhibitory effect on the NF-κB pathway by targeting TLR4. RV01 could be a potential protective agent in ischemic stroke treatment.

Eudesmane sesquiterpenoids with inhibitory effects on NO production from Artemisia princeps.

Five undescribed eudesmane methyl esters (1-5), three undescribed eudesmane-12,6-olides (6-8), and 21 known analogues (9-29) were isolated from the aerial part of Artemisia princeps Pamp. Their structures were established by detailed analysis of the NMR and HRESIMS data. The absolute configurations of 1-8 were determined based on single-crystal X-ray diffraction analysis and ECD calculations. Moreover, the inhibitory effects on LPS-induced NO production in BV-2 microglial cells of all the isolated compounds were assessed. Except for compounds 2, 4, 10, and 11, the others showed significant inhibitory activities, with IC50 values of 0.73-18.66 µM, wherein the potential structure-activity relationship was also discussed.

FOXO1 regulates bovine skeletal muscle cells differentiation by targeting MYH3.

The growth and development of bovine skeletal muscle and beef yield is closely intertwined. Our previous research found that forkhead box O1 (FOXO1) plays an important role in the regulation of beef muscle formation, but its specific mechanism is still unknown. In this study, we aimed to clarify the regulatory mechanism of FOXO1 in proliferation and differentiation of bovine skeletal muscle cells (BSMCs). The results showed that interfering with FOXO1 can promote proliferation and the cell G1/S phase of BSMCs by up-regulating the expression of PCNA, CDK1, CDK2, CCNA2, CCNB1, CCND1 and CCNE2. Besides, interfering with FOXO1 inhibited the apoptosis of BSMCs by up-regulating the expression of anti-apoptosis gene BCL2, while simultaneously down-regulating the expression of the pro-apoptosis genes BAD and BAX. Inversely, interfering with FOXO1 can promote the differentiation of BSMCs by up-regulating the expression of myogenic differentiation marker genes MYOD, MYOG, MYF5, MYF6 and MYHC. Furthermore, RNA-seq combined with western bolt, immunofluorescence and chromatin immunoprecipitation analysis showed that FOXO1 could regulate BSMCs differentiation process by influencing PI3K-Akt, Relaxin and TGF-beta signaling pathways, and target MYH3 for transcriptional inhibition. In conclusion, this study provides a basis for studying the role and molecular mechanism of FOXO1 in BSMCs.

Hysterolides A-I, dimeric or monomeric sesquiterpene lactones from Parthenium hysterophorus L.

Nine undescribed sesquiterpene lactones, including two pseudoguaianolide dimers (1 and 2), a pseudoguaiac dilactone (3), and six pseudoguaianolides (4-9), along with 13 known analogues (10-22) were isolated from Parthenium hysterophorus. Among them, hysterolide A (1) possesses an unusual carbon skeleton with a unique cyclobutane ring connecting two pseudoguaianolides. Hysterolide C (3) is a sesquiterpene dilactone incorporating a bicyclo[5.1.0]octane core. Spectroscopic analyses, 13C NMR and ECD calculations, and X-ray diffraction elucidated their structures and absolute configurations. Moreover, all the isolates were assayed for their anti-inflammatory activities by inhibiting LPS-induced nitric oxide production in BV-2 microglia cells, wherein, nine compounds displayed significant inhibitory activities with IC50 of 0.52-6.32 µM. Furthermore, the preliminary structure-activity relationship was also established.

Inhibition of TLR4 Signaling by Isorhapontigenin Targeting of the AHR Alleviates Cerebral Ischemia/Reperfusion Injury.

Ischemic stroke is a major risk factor in human health, yet there are no drugs to cure cerebral ischemia/reperfusion injury (CIRI). Inflammation plays a fundamental role in the consequences of CIRI. Isorhapontigenin (ISOR) exhibits great anti-inflammatory activity; however, it is unclear whether ISOR can treat ischemic stroke through an anti-inflammation effect. Here, middle cerebral artery occlusion/reperfusion (MCAO/R) was used to investigate the effects of ISOR on CIRI. The in vitro activity was measured in BV-2 cells exposed to oxygen-glucose deprivation/reperfusion. As measured by neurological scores, brain water content, and infarction, neurological dysfunction was improved in the ISOR group. The neuronal death and microglial activation in the ipsilateral cortex were reduced by ISOR. TLR4 signaling was significantly inhibited by ISOR in vivo and in vitro. By reverse molecular docking, cellular thermal shift, and drug affinity-responsive target stability assays, an aryl hydrocarbon receptor (AHR) was found to be a target of ISOR. Furthermore, AHR knockdown blocked the effect of ISOR on TLR4 signaling, suggesting that ISOR may regulate TLR4-mediated inflammation through AHR, thereby protecting neurons from CIRI. This study demonstrated that ISOR is a promising drug candidate for the treatment of ischemic stroke and provided a theoretical basis for the development of the medicinal value of ISOR-derived foods, such as grapes.

Pterostilbene participates in TLR4- mediated inflammatory response and autophagy-dependent Aß1-42 endocytosis in Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD), the most prevalent form of dementia, remains untreatable. One of the factors that contributes to its progression is microglia-mediated inflammation. Pterostilbene, a compound isolated from Chinese dragon's blood, can reduce inflammation caused by overactive microglia. However, its effects on AD transgenic animals and the possible underlying mechanism remain unknown. METHODS: We evaluated the effect of pterostilbene on learning and memory difficulties in transgenic APP/PS1 mice. We used immunofluorescence to detect microglial activation and Aß aggregation. We explored the cellular mechanism of pterostilbene by establishing LPS- stimulated BV2 cells and oAß1-42- exposed HEK 293T cells that overexpress TLR4 and/or MD2 via lentivirus. We applied flow cytometry and immunoprecipitation to examine how pterostilbene regulates TLR4 signaling. RESULTS: Pterostilbene enhanced the learning and memory abilities of APP/PS1 mice and reduced microglial activation and Aß aggregation in their hippocampus. Pterostilbene alleviated oAß1-42-induced inflammation, which required the involvement of MD2. Pterostilbene disrupted the binding between TLR4 and MD2, which may further prevent TLR4 dimerization and subsequent inflammatory response. Moreover, pterostilbene restored the impaired endocytosis of oAß1-42 through an autophagy-dependent mechanism. CONCLUSION: This is the first demonstration that pterostilbene can potentially treat AD by blocking the interaction of TLR4 and MD2, thereby suppressing TLR4-mediated inflammation.

Application of different proportions of sweet sorghum silage as a substitute for corn silage in dairy cows.

This experiment explored the effects of different proportions of sweet sorghum silage as a substitute for corn silage on dry matter intake (DMI), milk yield, milk quality, apparent digestibility, rumen fermentation parameters, serum amino acid profile, and rumen microbial composition of dairy cows. A total of 32 mid-lactation Holstein dairy cows with similar body weights and parities were randomly divided into four treatments: 100% corn silage +0% sorghum silage (CON), 75% corn silage +25% sorghum silage (CS1), 50% corn silage +50% sorghum silage (CS2), and 25% corn silage +75% sorghum silage (CS3). The milk yield was increased (linear, p = .048) as the proportion of sweet sorghum increased. Linear (p = .003) and quadratic (p = .046) increased effects were observed in milk fat as corn silage was replaced with sorghum silage. Compared with the CON diet group, the CS2 and CS3 diet groups had lower dry matter (DM) (linear, p < .001), ether extract (EE) (linear, p < .001), and gross energy (GE) (linear, p = .001) digestibility of the dairy cows. The ruminal fluid aspartate (Asp) level decreased (linear, p = .003) as the proportion of sweet sorghum increased. Linear (p < .05) and quadratic (p < .05) increased effects were observed for the contents of threonine (Thr), glycine (Gly), valine (Val), leucine (Leu), tyrosine (Tyr), and histidine (His) in rumen fluid with the replacement of corn silage with sorghum silage. Cows fed the CS3 diet had greater Faecalibacterium, Bacteroides, and Prevotella ruminicola content/copy number than those fed the CON diet (p < .05). In conclusion, feeding sorghum silage as a replacement for corn silage could increase the milk yield and fat, promote the growth of rumen microbes, and provide more rumen fluid amino acids for the body and microbial utilization. We believe that sorghum silage is feasible for dairy cows, and it is reasonable to replace corn silage with 75% sorghum silage.

Interaction of C/EBPß with SMAD2 and SMAD4 genes induces the formation of lipid droplets in bovine myoblasts.

As a key component of Transforming growth factor-ß (TGF-ß) pathway, Smad2 has many crucial roles in a variety of cellular processes, but it cannot bind DNA without complex formation with Smad4. In the present study, the molecular mechanism in the progress of myogenesis underlying transcriptional regulation of SMAD2 and SMAD4 had been clarified. The result showed the inhibition between SMAD2 and SMAD4, which promotes and inhibits bovine myoblast differentiation, respectively. Further, the characterization of promoter region of SMAD2 and SMAD4 was analyzed, and identified C/EBPß directly bound to the core region of both SMAD2 and SMAD4 genes promoter and stimulated the transcriptional activity. However, C/EBPß has lower expression in myoblasts which plays vital function in the transcriptional networks controlling adipogenesis, while the overexpression of C/EBPß gene in myoblasts significantly increased SMAD2 and SMAD4 gene expression, induced the formation of lipid droplet in bovine myoblasts, and promoted the expression of adipogenesis-specific genes. Collectively, our results showed that C/EBPß may play an important role in the trans-differentiation and dynamic equilibrium of myoblasts into adipocyte cells via promoting an increase in SMAD2 and SMAD4 gene levels. These results will provide an important basis for further understanding of the TGFß pathway and C/EBPß gene during myogenic differentiation.

Influence of Parturition on Rumen Bacteria and SCFAs in Holstein Cows Based on 16S rRNA Sequencing and Targeted Metabolomics.

The rumen fluids from ten cows at Day 3~5 before calving and Day 0 after calving were collected to analyze the composition and quantity of bacterial communities and concentrations of SCFAs. The results showed that the relative abundances of unidentified Lachnospiraceae, Acetitomaculum, Methanobrevibacter, Olsenella, Syntrophococcus, Lachnospira, and Lactobacillus genera were significant increased (p < 0.05), while that of unidentified-Prevotellaceae was notably decreased after calving (p < 0.05). In addition, the concentrations of acetic acid, propionic acid, butyric acid, and caproic acid obviously decreased after calving (p < 0.01). Our findings show that parturition altered the rumen microbiota and their fermentation ability in dairy cows. This study defines a rumen bacteria and metabolic profile of SCFAs associated with parturition in dairy cows.

Effects of Bacillus coagulans and Lactobacillus plantarum on the Fermentation Characteristics, Microbial Community, and Functional Shifts during Alfalfa Silage Fermentation.

This study aimed to investigate the potential of Bacillus coagulans (BC) as an inoculant in alfalfa silage fermentation. Fresh alfalfa was harvested at a dry matter (DM) content of 329.60 g/kg fresh weight (FW), and inoculated without (CON) or with BC (1 × 106 CFU/g FW), Lactobacillus plantarum (LP, 1 × 106 CFU/g FW), and their combinations (LP+BC, 1 × 106 CFU/g FW, respectively). Samples were taken at 3, 7, 14, 30, and 60 d, with three replicates for each. The prolonged ensiling period resulted in a decrease in pH values and an increase in lactic acid (LA) concentrations in alfalfa silages. After 60 d of fermentation, the application of BC and LP decreased the pH values and increased LA concentrations in treated silages, especially when their combination was applied. Application of BC preserved more water-soluble carbohydrates (WSC), and further application of BC increased WSC in LP+BC-treated silage compared to LP-treated silage. There was no significant difference in the crude protein (CP) content between the CON and treated silages, however, the BC and LP treatments reduced the ammonia nitrogen (NH3-N) concentration, especially when their combination was applied. Additionally, the BC and LP-treated silages had lower neutral detergent fiber (NDF) and acid detergent fiber (ADF) when compared to the CON silage (p < 0.001). Inoculants also increased Lactobacillus abundance and decreased Enterococcus abundance after 60 d of fermentation. Spearman's rank correlation analysis revealed a positive correlation between LA concentration and Lactobacillus abundance. It was noteworthy that LP, BC, and their combination increased the relative abundances of carbohydrate metabolism, energy metabolism, cofactors, and vitamin metabolism, decreasing the relative abundances of amino acid metabolism and drug resistance: antimicrobial. Therefore, the inclusion of BC increased the fermentation quality of alfalfa silage, with the optimal combination being LP+BC. According to the findings, BC could be considered a viable bioresource for improving fermentation quality.

Effect of endogenous sodium and potassium ions in plants on the quality of alfalfa silage and bacterial community stability during fermentation.

This study investigated the impact of endogenous sodium and potassium ions in plants on the quality of alfalfa silage, as well as the stability of bacterial communities during fermentation. Silage was produced from the fermented alfalfa, and the chemical composition, fermentation characteristics, and microbiome were analyzed to understand their interplay and impact on silage fermentation quality. The alfalfa was cultivated under salt stress with the following: (a) soil content of <1‰ (CK); (b) 1‰-2‰ (LP); (c) 2‰-3‰ (MP); (d) 3‰-4‰ (HP). The results revealed that the pH of silage was negatively correlated with the lactic acid content. With the increase of lactic acid (LA) content increased (26.3-51.0 g/kg DM), the pH value decreased (4.9-5.3). With the increase of salt stress, the content of Na+ in silage increased (2.2-5.4 g/kg DM). The presence of endogenous Na+ and K+ ions in plants significantly affected the quality of alfalfa silage and the dynamics of bacterial communities during fermentation. Increased salt stress led to changes in microbial composition, with Lactococcus and Pantoea showing a gradual increase in abundance, especially under high salt stress. Low pH inhibited the growth of certain bacterial genera, such as Pantoea and Pediococcus. The abundance of Escherichia-Shigella and Comamonas negatively correlated with crude protein (CP) content, while Enterococcus and Lactococcus exhibited a positive correlation. Furthermore, the accumulation of endogenous Na+ in alfalfa under salt stress suppressed bacterial proliferation, thereby reducing protein degradation during fermentation. The pH of the silage was high, and the LA content was also high. Silages from alfalfa under higher salt stress had higher Na+ content. The alpha diversity of bacterial communities in alfalfa silages showed distinct patterns. Desirable genera like Lactococcus and Lactobacillus predominated in silages produced from alfalfa under salt stress, resulting in better fermentation quality.

Dietary tea tree (Melaleuca alternifolia) oil supplementation enhances the expressions of amino acid transporters in goat ileal mucosa and improves intestinal immunity.

Tea tree oil (TTO) is a plant-derived additive with anti-inflammatory, bactericidal, and growth-promoting properties. However, little is known about the effects of TTO on intestinal amino acid transport and immune function in goats. Twenty-four Ganxi goats (initial body weight of 13.5 ± 0.70 kg) were randomly allotted two treatments and fed either control (CON) or CON+TTO (0.2 ml/kg) diet. The addition of TTO to the diet significantly decreased (p < .05) tumor necrosis factor-α content and increased (p < .05) interleukin-2 (IL-2) content in goat serum; significantly decreased (p < .05) IL-12, and increased (p < .05) IL-2 content in goat ileal mucosa; significantly increased (p < .05) secreted IgA content in the jejunal and ileal mucosa; significantly upregulated (p < .05) IL-2 and downregulated (p < .05) IL-12 at the mRNA level in the ileal mucosa; significantly elevated the levels of serine, arginine, and total amino acids in the ileal mucosa (p < .05); significantly upregulated (p < .05) SLC1A1 and SLC7A1 in the ileum; and significantly enhanced (p < .05) the protein expression of Claudin-1 in the ileal mucosa. In summary, adding 0.2 ml/kg of TTO to the diet enhanced SLC1A1 and SLC7A1 mRNA expression in the ileal mucosa, and SLC1A1 and SLC7A1 could transport serine and arginine from the chyme to the ileal mucosa. Thus, increased serine and arginine content in the mucosa could improve intestinal immunity. TTO supplementation upregulated the expression of IL-2 and Claudin-1 in goat ileal mucosa, and enhanced immune function in the intestine.

Dietary α-lipoic acid supplementation improves postmortem color stability of the lamb muscles through changing muscle fiber types and antioxidative status.

This study investigated the effect of dietary α-lipoic acid (600 mg/kg) supplementation on the postmortem color stability of the biceps femoris from lambs. The results showed that dietary α-lipoic acid supplementation increased a* and decreased b* and metmyoglobin (MMb) percentage of the biceps femoris with the time of storage (P < 0.05). The content of malondialdehyde (MDA) reduced with the time of storage after treatment with α-lipoic acid (P < 0.05). α-lipoic acid increased the myoglobin (Mb) content, and myosin heavy chain I (MyHC I) gene expression but decreased glycogen content, lactate dehydrogenase (LDH) activity, and MyHC IIb gene expression (P < 0.05). The T-AOC value, catalase (CAT) activity, and expression of SOD and CAT gene expression increased after α-lipoic acid treatment (P < 0.05). Therefore, dietary α-lipoic acid supplementation improved the meat color by regulating muscle fiber types and inhibited glycolysis. Moreover, α-lipoic acid maintained meat color stability by effectively inhibiting muscle oxidation via enhancing the antioxidant capacity.

Effects of Galactomannan Oligosaccharides on Growth Performance, Mycotoxin Detoxification, Serum Biochemistry, and Hematology of Goats Fed Mycotoxins-Contaminated Diets.

This study was conducted to investigate the protective effects of mycotoxin adsorbent galactomannan oligosaccharides (GMOS) on growth performance, fermentation parameters, mycotoxins residues, serum biochemistry and oxidative stress parameters of the goats. The in vitro test indicated that 0.05% GMOS outperformed yeast cell wall (YCW) and montmorillonite (MMT) in aflatoxins absorption. Then 20 3-month-old Xiangdong black goats (15.0 ± 1.9 kg) were randomly divided into two dietary treatments for the animal test. The control group (CON group) was fed a multi-mycotoxins contaminated diet, whereas the experimental group (GMOS group) received multi-mycotoxins contaminated diet plus 0.05% GMOS. The trail lasted for 60 days, with 12 days of adaptation period and 48 days of formal experiment period. There were no treatment effects (P > 0.10) on growth performance, serum antioxidant capacity and activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The concentrations of zearalenone in the rumen were lower (P < 0.05) in the GMOS group. GMOS significantly reduced (P < 0.05) propionate concentration in the cecum, resulting in a rise (P < 0.01) in acetate/propionate ratio in GMOS as compared to CON. Goats of GMOS exhibited considerably greater (P < 0.05) levels of creatine kinase but lower (P = 0.02) levels of creatinine than CON. Compared with CON, GMOS supplementation significantly increased (P < 0.05) platelet count (PLT), platelet volume distribution width (PDW), and platelet hematocrit (PCT), while decreased (P < 0.05) albumin content (ALB). The 0.05% GMOS protected goats in ruminal fermentation parameters, mycotoxins residues and serum biochemistry. Moreover, GMOS had no adverse effect on goat health. To our knowledge, this is the first report of GMOS in small ruminants. These findings suggested the feasibility of dietary GMOS as a health-maintaining addictive in goat diets.

Microbial Community, Fermentation Quality, and in vitro Degradability of Ensiling Caragana With Lactic Acid Bacteria and Rice Bran.

This study aimed to assess the effects of microbial inoculants and growth stage on fermentation quality, microbial community, and in vitro degradability of Caragana silage from different varieties. Caragana intermedia (CI) and Caragana korshinskii (CK) harvested at the budding (BU) and blooming (BL) stages were used as raw materials to prepare silage, respectively. The silages at each growth stage were treated for ensiling alone (control), with 5% rice bran (RB), a combination of RB with commercial Lactobacillus plantarum (RB + LP), and a combination of RB with a selected strain Lactobacillus plantarum L694 (RB + L694). The results showed that the crude protein (CP) content of CI was higher than that of CK, and delay in harvest resulted in greater CP content in Caragana at BL stage. After 60 days of fermentation, the concentrations of lactic acid (LA) in the RB + L694 treatments were higher than those in control treatments (p < 0.05), while the pH, concentrations of NH3-N, neutral detergent fiber with the addition of α-amylase (aNDF) were lower than those in control treatments (p < 0.05). RB + L694 treatments could decrease acid detergent fiber (ADF) content except in CIBL. In CK silages, adding RB + L694 could reduce bacterial diversity and richness (p < 0.05). Compared with the control, RB + L694 treatment contained higher Lactobacillus and Enterobacter (p < 0.05). In vitro NDF and DM degradability (IVNDFD and IVDMD) was mostly affected by growth period, and additive RB + l694 treatment had higher IVDMD and lower IVNDFD than other treatments (p < 0.05). Consequently, the varieties, growth stages, and additives could influence the fermentation process, while the blooming stage should be selected in both Caragana. Furthermore, the results showed that RB and L. plantarum could exert a positive effect on fermentation quality of Caragana silage by shifting bacterial community composition, and RB + L694 treatments outperformed other additives.

Tissue Expression Analysis, Cloning, and Characterization of the 5'-Regulatory Region of the Bovine LATS1 Gene.

As a member of the large tumor suppressor (LATS) gene family, LATS1 plays an important role in regulating muscle growth and development. In this study, we determined the distinct exhibit patterns of tissue expression of bovine LATS1. Further, we determined the functional proximal minimal promoter of bovine LATS1 and identified the key transcription factors in the core promoter region to elucidate its molecular regulation mechanism. The results showed that bovine LATS1 was highly expressed in the longissimus thoracis and upregulation in infancy muscle. An electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay in combination with site-directed mutation and small interfering RNA (siRNA) interference demonstrated that myogenic differentiation 1 (Myod1) and myocyte enhancer factor 2A (MEF2A) binding in the core promoter region (-298/-123 bp) play important roles in the transcriptional regulation of the bovine LATS1 promoter. Taken together, these interactions provide insight into the regulatory mechanisms of LATS1 transcription in mediating skeletal muscle growth in cattle.

Xanthones from the stems of Calophyllum membranaceum Gardn. et Champ. and their anti-inflammatory activity.

Eighteen undescribed xanthones, including six pairs of xanthone enantiomers, three xanthones, and three xanthone glycosides, together with one pair of known xanthone enantiomers and 12 known xanthones, were isolated from the stems of Calophyllum membranaceum Gardn. et Champ. Their structures were elucidated by spectroscopic analysis, and the absolute configuration of the enantiomers was determined by using experimental and calculated electronic circular dichroism data. All compounds were screened for their anti-inflammatory effects on LPS-induced BV-2 microglial cells. Among them, six compounds showed remarkable activities with IC50 values of 7.8-36.0 µM.

Integrated Metabolomics and Transcriptome Revealed the Effect of Fermented Lycium barbarum Residue Promoting Ovis aries Immunity.

Lycium barbarum residue contains abundant bioactive nutrients which can be used as feed supplement. The fermentation treatment of plant residue can promote the utilization of nutrients, rumen digestion, and the growth and immunity of animals. Based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) metabolomics and in-depth transcriptome analysis, the study tested the mechanisms of Lycium barbarum residue (RW) and fermented Lycium barbarum residue (RFW) on meat quality and immunity of sheep. Fifty-four Tan sheep were randomly divided into control, RFW or RW treatments. Data showed that RFW and RW increased the carcass weight, fat content, ash content and reduced the cooking loss of lamb. RFW performed more significant effects on activating immune-related genes than those of RW. The expression of chemokines and immune-related pathways, such as signaling pathways of interleukin-17 signaling pathway and NOD-like receptor signaling pathway, were elevated in sheep fed RFW. RW increased the diversity in rumen metabolites, especially compositions of lipids, organic acids and organ heterocyclic compounds. RFW affected numerous compounds which are closely correlated with the activation of immune genes. In conclusion, RFW could represent a valuable strategy to improve growth performance and immunity of sheep.

N-Carbamoylglutamate Supplementation on the Digestibility, Rumen Fermentation, Milk Quality, Antioxidant Parameters, and Metabolites of Jersey Cattle in High-Altitude Areas.

This study aimed to assess the impact of the dietary supplementation of N-carbamoylglutamate (NCG) on nutrient digestibility, rumen fermentation, milk quality, oxidative stress, and metabolites in the plasma and feces of Jersey cattle under high altitude with the hypoxic condition. A total of 14 healthy lactating Jersey dairy cows with similar body conditions were selected and randomly divided into 2 groups. The control group (CON group, N = 6 replicates) was fed with a conventional complete diet, whereas the experimental group (NCG group, N = 8 replicates) received 20 g/d per head NCG supplementation. The experiment lasted for 60 days, the adaptation period was 12 days, and the formal experiment period was 48 days. Except that the NCG group showed an upward trend in dry matter intake (DMI) (p = 0.09) and the fermentation parameters, the molar proportion of butyric acid tended to decrease (p = 0.08); the two groups had no significant differences (p > 0.05) in nutrients digestibility, plasma immunity, and antioxidant ability. However, compared with the CON group, the milk fat rate and blood oxygen saturation of the NCG group showed an upward trend (p = 0.09). For indexes associated with altitude stress, the contents of thyroxine, transferrin, and endothelin both decreased significantly (p < 0.05) in the NCG group. Meanwhile, heat shock protein (p = 0.07) and aldosterone (p = 0.06) also showed a downward trend. A total of 114 different metabolites were identified from feces and plasma, 42 metabolites were derived from plasma that mainly included 5 kinds of Super Class, and 72 metabolites were derived from feces that mainly included 9 kinds of Super Class. The significantly increased plasma differential metabolites were 2,5-dihydroxybenzoate and salicyluric acid, and the significantly increased fecal differential metabolites were Butenafine (fold change > 2). Pathway analysis showed that after applying NCG as a feed additive, the changes of the Jersey dairy cows mainly focused on amino acid metabolism and lipid metabolism. These results indicated that adding NCG to the diet can prevent the hypoxic stress state of lactating Jersey cows in high-altitude areas and has a tendency to improve milk quality.

Influence of Condensed and Hydrolysable Tannins on the Bacterial Community, Protein Degradation, and Fermentation Quality of Alfalfa Silage.

This study evaluated the effects of hydrolysable tannin (HT) and condensed tannin (CT) on the bacterial community, fermentation quality, and proteolysis of alfalfa silage. Alfalfa was wilted to a dry matter (DM) of 35% fresh weight and ensiled with or without 4% HT or 4% CT. The application rates of tannins were based on fresh weight, and each treatment was ensiled in triplicate. After 60 d of fermentation, the CT-treated group had lower concentrations of ammonia nitrogen (NH3-N) and free amino acid nitrogen (AA-N), but greater lactic acid concentration, than those in the control and HT-treated silage (p < 0.05). Compared to the control group, the application of tannins increased the abundance of Pseudomonas (negatively correlated with aminopeptidases activity), and decreased the abundance of Pediococcus­which was positively correlated with aminopeptidases activity­and the concentrations of non-protein nitrogen (NPN), NH3-N, and AA-N. The application of HT decreased the abundance of Lactobacillus and increased the abundances of Enterococcus, while the opposite results were observed in the CT-treated group. The application of HT and CT reduced the proteolysis in treated silages, but the two were different in terms of their mechanism and their effects on the bacterial communities of the alfalfa silage.