The effects of concurrent increases in supplementation of calcium and phytase on growth performance, balance of Ca and P, and bone mineralization in nursery pigs.

The objective of this study was to investigate the effects of concomitantly increasing supplementation of Ca and phytase on growth performance, balance of Ca and P, and bone mineralization in nursery pigs. There were eight experimental diets. The positive control (PC) one and two were formulated to contain 0.64% and 0.85% total Ca, respectively, whereas the dietary concentrations of other nutrients were identical and adequate. The negative control (NC) was deficient in total Ca (0.48%) and total P (0.41%). Five combinations of incremental levels of Ca and phytase (0.48% and 1,750 phytase units [FYT]/kg, 0.52% and 2,000 FYT/kg, 0.55% and 2,250 FYT/kg, 0.59% and 2,600 FYT/kg, and 0.63% and 3,000 FYT/kg) were added to the NC to establish the remaining five experimental diets. Each diet was fed to six pens of six pigs (three barrows and three gilts per pen). All diets contained 3 g/kg TiO2, and fecal samples were collected from each pen during the trial. In the end, one pig per pen was euthanized to collect the right tibia and urine in bladder. The results showed that the pigs of NC gained less weight, consumed less feed, and utilized feed less efficiently than their counterparts fed the PC and the treatments with phytase (P < 0.01). With increasing supplementation of Ca and phytase, there was a tendency for gain:feed to decrease (P < 0.10). There was a significant reduction in bone dry weight; and in percentages, as well as weights of bone ash, Ca, and P; in pigs of NC compared with pigs of PC1, PC2, or phytase treatments. In comparison to PC2, PC1 and phytase treatments resulted in a higher percentage of bone P and greater weights of bone ash, Ca, and P (P < 0.05). There was no significant effect of concurrent supplementation of Ca and phytase on bone mineralization. The NC had significantly lower apparent total tract digestibility (ATTD) of Ca and P, lower concentrations of digestible Ca and P, but a higher ATTD Ca/ATTD P ratio than PC1, PC2, or the phytase treatments. The averages of ATTD of Ca and P in treatments with phytase were significantly higher than PC1 or PC2 (P < 0.01). With increasing addition of Ca and phytase, the ATTD of P, digestible Ca and P, and the ATTD Ca/ATTD P ratio increased linearly (P < 0.05), which contrasted with a linear reduction in ATTD of Ca (P < 0.05). Meanwhile, there was a linear (P < 0.01) increase in the concentration of urinary Ca. In conclusion, increasing the dietary supplementation of phytase in conjunction with the increasing dietary Ca level increased the dietary ATTD Ca/ATTD P ratio without damaging the absorption of P in the current study. The higher ATTD Ca/ATTD P ratio did not improve the bone mineralization markedly and thus the extra Ca was voided through urine.

C-X-C motif chemokine ligand 12 improves the developmental potential of bovine oocytes by activating SH2 domain-containing tyrosine phosphatase 2 during maturation†.

In vitro maturation of mammalian oocytes is an important means in assisted reproductive technology. Most bovine immature oocytes complete nuclear maturation, but less than half develop to the blastocyst stage after fertilization. Thus, inefficient in vitro production is mainly caused by a suboptimal in vitro culture process, in which oocyte quality appears to be the limiting factor. In our study, a potential maternal regulator, C-X-C motif chemokine ligand 12, was identified by analyzing transcriptome data. C-X-C motif chemokine ligand 12 supplementation promoted the developmental potential of oocytes by improving protein synthesis and reorganizing cortical granules and mitochondria during in vitro maturation, which eventually increased blastocyst formation efficiency and cell number after parthenogenesis, fertilization, and cloning. All these promoting effects by C-X-C motif chemokine ligand 12 were achieved by activating SH2 domain-containing tyrosine phosphatase 2, thereby promoting the mitogen-activated protein kinase signaling pathway. These findings provide an in vitro maturation system that closely resembles the maternal environment to provide high-quality oocytes for in vitro production.

The effects of increasing dietary total Ca/total P ratios on growth performance, Ca and P balance, and bone mineralization in nursery pigs fed diets supplemented with phytase.

The objective of this study was to investigate the effects of increasing dietary total Ca/total P ratios on growth performance, digestibility of Ca and P, bone mineralization, and concentrations of Ca and P in urine and plasma in nursery pigs. There were six diets in a randomized complete block design, including one positive control and five diets corresponding to five total Ca/total P ratios: 0.55, 0.73, 0.90, 1.07, and 1.24 (analyzed as 0.58, 0.75, 0.93, 1.11, and 1.30). These five diets were deficient in P but supplemented with 1,000 phytase units/kg feed. Each diet was fed to six pens of eight pigs (four barrows and four gilts per pen). All diets contained 3 g/kg TiO2, and fecal samples were collected from each pen on days 5-7 of trial. At the end, one pig per pen was sacrificed to collect the right tibia and urine in the bladder. The results showed that increasing dietary Ca/P ratio to 0.93 increased gain:feed but then gain:feed decreased as the Ca/P ratio was increased to 1.30 (linear and quadratic, P < 0.05). Although average daily gain and final BW were unaffected by changing Ca/P ratio in diet, dry bone weight; weights of bone ash, Ca and P; and bone Ca/P ratio increased linearly (P < 0.001) with increasing dietary Ca/P ratio. The percent bone Ca showed a tendency to increase (P = 0.064). Increasing dietary Ca/P ratio decreased apparent total tract digestibility (ATTD) of Ca and P linearly (P < 0.05) and the concentration of digestible P linearly (P < 0.001), but increased the concentration of digestible Ca (linear and quadratic effects: P < 0.01) and the digestible Ca/P ratio (linear effect: P < 0.001). In plasma, the concentration of Ca increased both linearly (P < 0.01) and quadratically (P = 0.051), whereas the concentration of P tended (linear and quadratic, P < 0.10) to decrease with increasing dietary Ca/P ratio. Similarly, in urine, the concentration of Ca increased both linearly and quadratically (P < 0.05), whereas the concentration of P decreased linearly (P < 0.01). In conclusion, increasing the dietary Ca/P ratio reduced feed efficiency but increased bone mass and the amounts of Ca and P deposited in bone of nursery pigs fed diets supplemented with 1,000 FYT/kg phytase. The increases in bone growth led to a reduction of urinary P excretion that exceeded the decreased digestible P supplied in diet with the widening dietary Ca/P ratios.

Use of fixed calcium to phosphorus ratios in experimental diets may create bias in phytase efficacy responses in swine.

The objective of this study was to investigate the effects of two dietary total Ca/P ratios on available P release by phytase, measured using growth performance and bone mineralization with 528 barrows and gilts according to a randomized complete block design. Three were 11 diets in a factorial of 2 by 4 plus 3, including 3 reference diets consisting of 0.25% (control), 0.70%, or 1.15% monocalcium phosphate (MCP) and 8 diets from combining 4 phytase doses (500, 1,000, 2,000, and 3,000 FYT/kg) with 0.25% MCP and 2 dietary Ca/P ratios (1.05 and 1.20). Each diet was fed to 6 pens of 8 pigs. All diets contained 3 g/kg TiO2, and fecal samples were collected from each pen on d 13-15 of trial. At the end of trial, one pig per pen was sacrificed to collect a tibia and urine in the bladder. The results showed that MCP improved growth performance linearly (P < 0.01), whereas both a linear and quadratic response was observed with the addition of phytase. The MCP increased the percent bone ash and weights of bone ash, Ca, and P linearly (P < 0.01). At both Ca/P ratios, increasing supplementation of phytase increased the percent bone ash and weights of bone ash, Ca, and P both linearly and quadratically (P < 0.05). Both MCP and phytase significantly increased digestibility of Ca and P as well as digestible Ca and P in diets and reduced the digestible Ca/P ratio. The dietary Ca/P ratio of 1.20 resulted in poorer feed utilization efficiency, more digestible Ca, greater percent bone ash, Ca, and P and heavier weights of bone Ca and P than the ratio of 1.05 (P < 0.05). The ratio of 1.20 elicited numerically higher available P release values from phytase, with percent bone ash and bone P weight as the response variables, but significantly lower values with gain:feed. The urinary concentration of Ca increased linearly (P < 0.01) with increasing digestible Ca/P ratios whilst urinary concentration of P decreased quadratically (P < 0.01). In conclusion, fixing the same total Ca/total P ratio in diets supplemented with increasing phytase dosing created an imbalance of digestible Ca and P, which could have an adverse effect on bone mineralization and thus compromise the phytase efficacy relative to mineral P.

Integrated RNA-Seq Analysis Uncovers the Potential Mechanism of the "Kidney Governing Bones" Theory of TCM.

Background: As in philosophy of traditional Chinese medicine (TCM), the theory of "kidney governing bones" has been demonstrated by a series of scientific studies. Furthermore, many groups including ours have explored the molecular mechanisms related to bone development, growth, and regeneration using modern biology technologies, such as RNA sequencing (RNA-Seq) and isobaric tags for relative and absolute quantification (ITRAQ), and have demonstrated that the underlying molecular mechanisms were highly consistent with the "kidney governing bones" theory. Objective: Kidney-yang deficiency (YD), as a pathological condition, has a passive effect on the skeleton growth; more specifically, it is a state of skeletal metabolic disorder. However, the exact molecular mechanisms related to the "kidney governing bones" theory under the control of multiple organs and systems are still unknown. Methods: In this study, we performed RNA-Seq analysis to investigate and compare the gene expression patterns of six types of tissue (bone, cartilage, kidney, testicle, thyroid gland, and adrenal gland) from YD rats and normal rats and analyzed the interaction effects controlled by multiple functional genes and signaling pathways between those tissues. Results: Our results showed that, in the state of YD, the functions of bone and cartilage were inhibited. Furthermore, multiple organs involving the reproductive, endocrine, and urinary systems were also investigated, and our results showed that YD could cause dysfunctions of these systems by downregulating multiple functional genes and signaling pathways that positively regulate the homeostasis of these tissues. Conclusion: We ensure that "kidney governing bones" was not a simple change in a single gene but the changes in complex biological networks caused by functional changes in multiple genes. This also coincides with the holistic view of TCM, which holds that the human body itself is an organic whole and the functional activities of each organ coordinate with each other.

Coenzyme Q10 Supplement Rescues Postovulatory Oocyte Aging by Regulating SIRT4 Expression.

BACKGROUND: High-quality of the oocyte is crucial for embryo development and the success of human-assisted reproduction. The postovulatory aged oocytes lose developmental competence with mitochondrial dysfunction and oxidative stress. Coenzyme Q10 (CoQ10) is widely distributed in the membranes of cells and has an important role in the mitochondrial respiration chain against oxidative stress and modulation of gene expression. OBJECTIVE: The objective of this study is to investigate the functions and mechanisms of CoQ10 on delaying postovulatory oocyte aging. METHODS: Quantitative real-time PCR and Immunofluorescence staining were used to determine the expression patterns of the biogenesis genes of CoQ10 in postovulatory aged oocytes compared with fresh oocytes. The mitochondrial function, apoptosis, reactive oxygen species (ROS) accumulation and spindle abnormalities were investigated after treatment with 10 µM CoQ10 in aged groups. SIRT4 siRNA or capped RNA was injected into oocytes to investigate the function of SIRT4 on postovulatory oocyte aging and the relationship between CoQ10 and SIRT4. RESULTS: Multiple CoQ10 biosynthesis enzymes are insufficient, and a supplement of CoQ10 can improve oocyte quality and elevate the development competency of postovulatory aged oocytes. CoQ10 can attenuate the aging-induced abnormalities, including mitochondrial dysfunction, ROS accumulation, spindle abnormalities, and apoptosis in postovulatory aged oocytes. Furthermore, SIRT4, which was first found to be up-regulated in postovulatory aged oocytes, decreased following CoQ10 treatment. Finally, knockdown of SIRT4 can rescue aging-induced dysfunction of mitochondria, and the efficiency of CoQ10 rescuing dysfunction of mitochondria can be weakened by SIRT4 overexpression. CONCLUSION: Supplement of CoQ10 protects oocytes from postovulatory aging by inhibiting SIRT4 increase.

The effect of short-term phytase supplementation on the apparent total tract digestibility of calcium and phosphorus and the reproductive performance of late gestation and lactating sows fed diets without mineral phosphorus.

The objective of this study was to measure apparent total tract digestibility (ATTD) of Ca and P as well as reproductive performance in late gestation and lactating sows supplemented with a novel phytase and to compare the response to phytase supplementation between late gestation and lactating sows. A total of 45 late gestation sows and 45 lactating sows were used in experiments 1 and 2, respectively, in a completely randomized design. The sows were provided with a control diet or the control diet supplemented with 187.5 or 375 FYT phytase/kg feed for 10 days. The diets were prepared according to the formulas in use for production but without any inorganic P supplement. Titanium dioxide was included at 3 g/kg feed as an indigestible marker. Each dietary treatment was replicated with 15 sows individually housed in farrowing stalls. The sows were allowed to adapt to the experimental diets for 5 days before a 5-d fecal collection by grab sampling, and the performance of the sows and their litters were measured until weaning. The results showed that the ATTD of Ca increased linearly (P < 0.001), while the ATTD of P increased both linearly and quadratically (P < 0.01) with increasing supplementation of phytase in both late gestation and lactating sows. There was no significant effect of phytase on the ATTD of dry matter, crude protein, and gross energy, and the performance of the sows and their progenies. The phytase added at 187.5 and 375 FYT/kg feed released 0.07% and 0.10% digested P, respectively, in late gestation sows, which compared with 0.09% and 0.12% digested P in lactating sows. In conclusion, a novel phytase at 187.5-375 FYT/kg feed could release 0.07-0.12% digestible P for sows. It appeared that using the P digestibility values of feed ingredients listed by NRC to formulate a diet for sows might overestimate dietary P supply and a greater response to phytase supplementation could be expected in lactating sows than in late gestation sows.

A new MNF-BM4D denoising algorithm based on guided filtering for hyperspectral images.

This paper proposed a new MNF-BM4D denoising algorithm based on guided filtering to improve the denoising performance of the state-of-the-art Block-Matching and 4D filtering(BM4D) algorithm for hyperspectral images in the spatial and spectral domain. BM4D is firstly used to denoise hyperspectral images. Then Minimum Noise Fraction(MNF) algorithm is introduced to distinguish between the main component and the noisy component. Finally, the guided image filtering technology is utilized to further improve the denoising performance. A number of experiments on both simulated and real data are conducted to validate the effective denoising performance of the proposed method. Therefore, the proposed algorithm can be considered as a promising technique for hyperspectral imagery denoising.

Phenotypic and Genetic Diversity of the Culinary-Medicinal Winter Mushroom Flammulina velutipes (Agaricomycetes) in China.

Flammulina velutipes is one of the mushrooms produced most widely in East Asia. In this study we investigated phenotypic variations of 7 agronomic traits in 37 F. velutipes strains from China, and we analyzed their genetic diversity with 70 simple sequence repeat markers. The Shannon information index and gene diversity were 0.894 and 0.478, respectively, demonstrating high genetic variation among the tested strains. Poor genetic variation was found among white strains, in contrast to yellow ones. Analysis of population structure resolved 2 unambiguous genetic groups in the tested F. velutipes strains, with little differentiation between them (FST = 0.016). Yellow cultivars possibly originated from indigenous wild strains in southwest China. Phenotypic correlations were identified among the 7 traits. In particular, stipe length (SL) was significantly positively correlated with yield, indicating that SL could be used as an index for breeding high-yield strains. The 2 genetic groups, and white strains and yellow ones, showed significant differences between SL, yield, and the time interval (days) from mycelial scratch to formation of the first fruiting body. These results indicate that these 3 traits were stratified by population structure. Detection of genetic and phenotypic variations would lay the groundwork for further breeding of elite F. velutipes strains.

Enhanced healing of rat calvarial critical size defect with selenium-doped lamellar biocomposites.

A 3D porous lamellar selenium-containing nano-hydroxyapatite (SeHAN)/chitosan (CS) biocomposite was synthesized. The selenium-containing hydroxyapatite (HA) grains of 150~200 nm in length and 20~30 nm in width were observed by dynamic light scattering and transmission electron microscopy. A combination of X-ray diffraction, Fourier-transform infrared spectroscopy, and SEM indicated that HA particles were uniformly dispersed in chitosan matrix and there was a chemical interaction between chitosan and HA. Then, a standard critical size calvarial bone defect was created in Wistar rats. In group 1, no implant was made in the defect. In groups 2 and 3, HA nanoparticles (HAN)/CS biocomposite and SeHAN/CS biocomposite were implanted into the defect, respectively. After 4 weeks, the histological assessment clearly exhibited no significant changes, only found some living cells anchored in the periphery of the implants. After 8 and 12 weeks, most newly formed osteoid tissue was found in the SeHAN/CS implant group. Additionally, the newly formed osteoid tissue, both at the edge and in the center of implants, was bioactive and neovascularized. Microfocus computerized tomography measurements also confirmed the much better quality of the newly formed bone tissue in SeHAN/CS implant group than that in HAN/CS implant group (p < 0.01). Collectively, the SeHAN/CS biocomposite, as a bioactive bone grafting substitute, significantly enhanced the repair of bone defect.