Characteristics of recombinant xylanase from camel rumen metagenome and its effects on wheat bran hydrolysis.

In the present study, we explored the effects of a novel xylanase from camel rumen metagenome (CrXyn) on wheat bran hydrolysis. CrXyn was heterologously expressed in Escherichia coli and showed maximum activity at 40 °C and pH 7.0. Furthermore, CrXyn exhibited preferential hydrolysis of xylan, but no obvious activity toward other substrates, including carboxymethylcellulose and Avicel. Using wheat straw xylan as a substrate, the Km and Vmax values for CrXyn were 5.98 g/L and 179.9 µmol xylose/min/mg protein, respectively. Mn2+ was a strong accelerator and significantly enhanced CrXyn activity. However, CrXyn activity was inhibited (~50 %) by 1 mM and 5 mM ethylenediaminetetraacetic acid (EDTA) and completely inactivated by 5 mM Cu2+. CrXyn tolerated 5 mM sodium dodecyl sulphate (SDS) and 15 % methanol, ethanol, and dimethyl sulfoxide (DMSO), with >50 % residual activity. CrXyn effectively hydrolyzed wheat bran, with xylobiose and xylotetraose accounting for 79.1 % of total sugars produced. A remarkable synergistic effect was found between CrXyn and protease, leading to an obvious increase in amino acids released from wheat bran compared with the control. CrXyn also enhanced the in vitro hydrolysis of wheat bran. Thus, CrXyn exhibits great potential as a feed additive to improve the utilization of wheat bran in monogastric animal production.

Characterization of yeast cell surface displayed Lentinula edodes xylanase and its effects on the hydrolysis of wheat.

The current study displayed a xylanase from Lentinula edodes on the surface of Pichia pastoris (sdLeXyn) and investigated its properties and effects on the wheat hydrolysis. Fluorescence microscope results showed that sdLeXyn was successfully anchored and displayed on the surface of P. pastoris X-33 cells. The highest activity of sdLeXyn was obtained at pH 3.0 and 50 °C. The sdLeXyn exhibited anti-high temperature property and showed broad temperature adaptability (>55% of the highest activity at 20-80 °C). The sdLeXyn was very stable at room temperature and could remain functionally stable at 50 °C for 3 h. The Km value was greater in sdLeXyn than that in free recombinant L. edodes xylanase. The sdLeXyn exhibited well resistance to Mn2+, Zn2+, Ca2+, Na+, Cu2+, Mg2+, K+, Ni2+ (1 mM and 5 mM) except Cu2+, which reduced the sdLeXyn activity by 54.5% at 5 mM dosage. The activity of sdLeXyn was increased by 42.6% by 5 mM Mn2+, 5 mM DTT, Trition X-100, and Tween 20 did not affect the activity of sdLeXyn, but SDS and EDTA slightly reduced it by 12.8% and 14.6%, respectively. The sdLeXyn could resist the degradation of pepsin, efficiently hydrolyzed wheat and reduced the viscosity of wheat hydrolysate. Current data indicate that the sdLeXyn has a potential as a feed additive to improve the utilization of wheat in poultry production.

Niacin mitigates rumen epithelial damage in vivo by inhibiting rumen epithelial cell apoptosis on a high concentrate diet.

To investigate the effects of niacin on rumen fermentation, rumen epithelial antioxidant activity, and rumen epithelial cell apoptosis on high concentrate (HC) diets, nine male Hu sheep were randomly fed one of three diets: low concentrate diet (LC; concentrate: forage (C:F) = 20:80, high concentrate diet (HC; C:F = 80:20), and HCN diet (HC diet + niacin at 800 mg/kg diet air-dry matter). Compared with the LC group, the HC group had a lower rumen pH, higher volatile fatty acids and lactic acid in the rumen, reduced activity of antioxidant enzymes and total antioxidant capacity, and increased malondialdehyde content in the rumen epithelium (P < 0.05). Rumen epithelial papilla morphology was decreased, and apoptosis-related indicators and serum inflammatory cytokines were increased in the HC group over the LC group (P < 0.05). Compared with the HC diet, the HCN diet increased rumen pH, rumen epithelium antioxidant capacity, and rumen epithelial papilla morphology, decreased rumen lactate content, serum inflammatory cytokines, and apoptosis-related indicators (P < 0.05). Therefore, adding 800 mg/kg niacin helped protect against rumen epithelial damage by avoiding drastic changes in the rumen environment and improved rumen epithelial antioxidant capacity to inhibit rumen epithelial cell apoptosis in sheep on a HC diet.

Dynamic mechanism of multiple bursting patterns in a whole-cell multiscale model with calcium oscillations.

The dynamic mechanism of a whole-cell model containing electrical signalling and two-compartment Ca2+ signalling in gonadotrophs is investigated. The transition from spiking to bursting by Hopf bifurcation of the fast subsystem about the slow variable is detected via the suitable parameters. When the timescale of K+ gating variable is changed, the relaxation oscillation with locally small fluctuation, chaotic bursting and mixed-mode bursting (MMB) are revealed through chaos. In addition, the bifurcation of [Ca2+]i with regard to [IP3] is analysed, showing periodic solutions, torus, period doubling solutions and chaos. Finally, hyperpolarizations and torus canard-like behaviours of the full system under a set of specific parameters are elucidated.

Global stability of age-of-infection multiscale HCV model with therapy.

In order to treat the diseases caused by hepatitis C virus (HCV) more efficiently, the concentration of HCV in blood, cells, tissues and the body has attracted widespread attention from related scholars. This paper studies a dynamic dependent HCV model (more specifically, including age structure and treatment methods model) that concludes states of infection-free and infected equilibrium. Through eigenvalue analysis and Volterra integral formula, it proves that $E_0$ is globally asymptotically stable when $\mathcal{R}<1$. After explaining the existence, uniqueness and positive properties of the solution of the system, we have proved the global asymptotic stability of $E^*$ when $\mathcal{R}>1$ by constructing a suitable Lyapunov function. Through the above proofs, it can be concluded that effective treatment measures can significantly reduce the number of HCVs, so many related researchers are aware of the importance of highly efficient nursing methods and are committed to applying relevant methods to practice.

Characteristics of a recombinant Fusarium verticillioides cutinase and its effects on enzymatic hydrolysis of rice straw.

The current study heterologously expressed a cutinase from Fusarium verticillioides by Pichia pastoris and investigated its properties and effects on the hydrolysis of rice straw. The optimal pH and temperature for F. verticillioides cutinase were 8.0 and 50 °C, respectively. F. verticillioides cutinase had poor thermal stability and could be inhibited by some metal ions, inhibitors, and detergents (5 mM), including Ni2+, Zn2+, Cu2+, Ca2+, Mn2+, sodium dodecyl sulfate, EDTA, and Tween-20. F. verticillioides cutinase could tolerate 15% methanol and dimethyl sulfoxide but was significantly repressed by 15% ethanol and acetone with 48% and 63% residual activity, respectively. F. verticillioides cutinase could degrade the cuticle of rice straw with palmitic acid and stearic acid as the main products. However, the dissolving sugars released from the rice straw treated with F. verticillioides cutinase were significantly reduced by 29.2 µg/mL compared with the control (107.9 µg/mL). Similarly, the reducing sugars produced from the cellulase hydrolysis of rice straw pretreated with F. verticillioides cutinase were reduced by 63.5 µg/mL relative to the control (253.6 µg/mL). Scanning electron microscopy results showed that numerous tuberculate or warty protrusions were present nearly everywhere on the surface of rice straw treated with F. verticillioides cutinase, and some protrusions even covered and blocked the stomata of the rice straw surface. Current limited data indicate that F. verticillioides cutinase might not be an appropriate choice for improving the utilization of agricultural straws.

Peptidoglycan derived from Lactobacillus rhamnosus MLGA up-regulates the expression of chicken ß-defensin 9 without triggering an inflammatory response.

Defensins are critical components of the innate immune system and play an important role in the integration of innate and adaptive immune responses. Although information on the immunomodulatory properties of peptidoglycan from bacteria is abundant, little is known about the ß-defensin induction effect of peptidoglycan from the probiotic Lactobacillus. This study investigated the effect of intact peptidoglycan from L. rhamnosus MLGA on the induction of avian ß-defensin 9 in chicken immune cells and intestinal explants. Peptidoglycan from Lactobacillus rhamnosus MLGA dose dependently promoted avian ß-defensin 9 mRNA expression in chicken PBMCs, splenocytes, thymocytes, hepatocytes, and chicken embryo jejunum, ileum, and cecum explants and increased the capacity of PBMC or splenocyte lysates to inhibit the growth of Salmonella Enteritidis. In contrast to the effect of L. rhamnosus MLGA-derived peptidoglycan, peptidoglycan derived from pathogenic Staphylococcus aureus reduced avian ß-defensin 9 mRNA expression in chicken PBMCs and splenocytes. The inducible effect of peptidoglycan from L. rhamnosus MLGA on avian ß-defensin 9 expression in PBMCs and splenocytes was observed without activation of the expression of associated pro-inflammatory cytokines IL-1ß, IL-8, and IL-12p40, whereas these cytokine expressions were suppressed by peptidoglycan hydrolysate obtained by lysozyme digestion. The results of the present study show the capability of peptidoglycan derived from L. rhamnosus MLGA to induce the antimicrobial peptide defensin while simultaneously avoiding the deleterious risks of an inflammatory response.

Niacin Protects against Butyrate-Induced Apoptosis in Rumen Epithelial Cells.

The effects and underlying mechanisms of butyrate and butyrate+niacin on apoptosis in sheep rumen epithelial cells were investigated. Cells were exposed to butyrate (0-140 mM) for 6 h. A low concentration (20 mM) of butyrate increased cell viability and promoted growth whereas high concentrations (40-140 mM) inhibited proliferation. Cells were then cocultured with 120 mM butyrate and niacin (0-100 mM) for 6 h. Niacin addition attenuated butyrate-induced cellular damage and promoted proliferation at 20-80 mM; 40 mM presented the optimal effect. Higher concentrations (100 mM) of niacin resulted in low cell viability. Subsequent experiments confirmed that 120 mM butyrate increased intracellular reactive oxygen species (ROS) production and reduced the intracellular total antioxidant capacity (T-AOC) versus the untreated control. Compared with 120 mM butyrate, cotreatment with 40 mM niacin significantly reduced the intracellular ROS content and increased the intracellular T-AOC. Flow cytometry analysis revealed that 120 mM butyrate increased the proportion of apoptotic cells by 17.8% versus the untreated control, and 120 mM butyrate+40 mM niacin treatment reduced the proportion of apoptotic cells by 28.6% and 39.4% versus the untreated control and butyrate treatment, respectively. Treatment with 120 mM butyrate increased caspase-9 and p53 mRNA levels and decreased the expression of Bcl-2 and Bax, and the Bcl-2/Bax ratio versus the untreated control. Treatment with 120 mM butyrate+40 mM niacin downregulated the expression of caspase-3 and p53 and increased the expression of Bcl-2 and Bax versus butyrate treatment alone but had no effect on the Bcl-2/Bax ratio. Thus, high concentrations of butyrate may induce rumen epithelial cell apoptosis by increasing oxidative stress and inducing caspase-9 and p53 expression. Cotreatment with niacin regulates apoptosis-related gene expression by reducing intracellular ROS production and DNA damage and downregulating caspase-3 and p53 expressions to protect rumen epithelial cells against butyrate-induced apoptosis.

Niacin alters the ruminal microbial composition of cattle under high-concentrate condition.

To understand the effects of niacin on the ruminal microbial ecology of cattle under high-concentrate diet condition, Illumina MiSeq sequencing technology was used. Three cattle with rumen cannula were used in a 3 × 3 Latin-square design trial. Three diets were fed to these cattle during 3 periods for 3 days, respectively: high-forage diet (HF; forage-to-concentrate ratio = 80:20), high-concentrate diet (HC; forage-to-concentrate ratio = 20:80), and HC supplemented with 800 mg/kg niacin (HCN). Ruminal pH was measured before feeding and every 2 h after initiating feeding. Ruminal fluid was sampled at the end of each period for microbial DNA extraction. Overall, our findings revealed that subacute ruminal acidosis (SARA) was induced and the α-diversity of ruminal bacterial community decreased in the cattle of HC group. Adding niacin in HC could relieve the symptoms of SARA in the cattle but the ruminal pH value and the Shannon index of ruminal bacterial community of HCN group were still lower than those of HF group. Whatever the diet was, the ruminal bacterial community of cattle was dominated by Bacteroidetes, Firmicutes and Proteobacteria. High-concentrate diet significantly increased the abundance of Prevotella, and decreased the abundance of Paraprevotella, Sporobacter, Ruminococcus and Treponema than HF. Compared with HC, HCN had a trend to decrease the percentage of Prevotella, and to increase the abundance of Succiniclasticum, Acetivibrio and Treponema. Increasing concentrate ratio could decrease ruminal pH value, and change the ruminal microbial composition. Adding niacin in HC could increase the ruminal pH value, alter the ruminal microbial composition.