Proteome changes of sheep rumen epithelium during postnatal development.

Background: The development of the rumen epithelium is a critical physiological challenge for sheep. However, the molecular mechanism underlying postnatal rumen development in sheep remains rarely understood. Results: Here, we used a shotgun approach and bioinformatics analyses to investigate and compare proteomic profiles of sheep rumen epithelium tissue on day 0, 15, 30, 45, and 60 of age. A total of 4,523 proteins were identified, in which we found 852, 342, 164, and 95 differentially expressed proteins (DEPs) between day 0 and day 15, between day 15 and day 30, between day 30 and day 45, between day 45 and day 60, respectively. Furthermore, subcellular localization analysis showed that the DEPs were majorly localized in mitochondrion between day 0 and day 15, after which nucleus proteins were the most DEPs. Finally, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that DEPs significantly enriched in mitochondrion, ubiquitination, histone modifications, glutathione synthase activity, and wnt and nortch signaling pathways. Conclusion: Our data indicate that the biogenesis of mitochondrion in rumen epithelial cell is essential for the initiation of rumen epithelial development. Glutathione, wnt signaling pathway and nortch signaling pathway participated in rumen epithelial growth. Ubiquitination, post-translational modifications of histone might be key molecular functions in regulating rumen epithelial development.

Ultrasensitive cascaded in-line Fabry-Perot refractometers based on a C-shaped fiber and the Vernier effect.

We propose and experimentally demonstrate a fiber refractometer based on a C-shaped fiber and the Vernier effect. The sensor is fabricated by cascading a single mode fiber (SMF) pigtail together with a C-shaped fiber segment and another SMF segment. Thus, the C-shaped fiber would constitute an open cavity (sensing cavity) in which test analytes could be filled, while the SMF segment would constitute another reference cavity. Due to the similar optical path length of these two cavities, the Vernier effect would be activated, thus forming spectral envelops in the reflection spectrum of the sensor. Variations in the refractive index (RI) of analytes would result in the shifts of the spectral envelops. Both theoretical calculations and experiments are carried out in the characterization of the sensor measuring liquid and gaseous analytes. The experimental sensitivity of the sensor is found to be ∼37238 nm/RIU for gas RI measurement. The proposed sensor features the advantages such as ease of fabrication, extremely high sensitivity, capability of sensing of both gaseous and liquid analytes, small footprint, and good mechanical strength. Compared to other existing Vernier effect-based fiber refractometers typically fabricated using PCFs, the proposed sensor would allow analytes to have much easier and quicker access to the sensor probe.

Proteomic and parallel reaction monitoring approaches to evaluate biomarkers of mutton tenderness.

Intensive fattening usually results in the changes of meat quality. Tenderness is a central attribute for mutton sensory qualities and consumers' choice. Here, we reported that intensive fattening mutton was more tender than that of traditionally raised sheep. By proteomic approach, we found 49 differentially expressed proteins in longissimus dorsi muscle. After bioinformatics analysis, 5 cytoskeletal proteins, 3 protein binding proteins and 7 metabolic enzymes were identified as potential biomarkers for mutton tenderness. Finally, we verified the expression of these abundant proteins by parallel reaction monitoring (PRM). Collectively, our results reveal that the mutton of sheep raised by intensive fattening is more tender than that of traditionally raised sheep. Myosin-2, myosin-13, vimentin, carbonic anhydrase, carbonic anhydrase-2, Glutathione S-transferase and Microtubule-associated protein 4 isoform X1 can be candidate biomarkers for mutton tenderness. Our data also indicate a central role of cytoskeletal proteins and metabolic enzymes in determining mutton tenderness.

Proteome changes of dairy calves rumen epithelium from birth to postweaning.

Background: Rumen epithelium plays a central role in absorbing, transporting, and metabolizing of short-chain fatty acids. For dairy calves, the growth of rumen papillae greatly enhances the rumen surface area to absorb nutrients. However, the molecular mechanism underlying dairy calves rumen postnatal development remains rarely understood. Results: Here, we firstly describe the histological change of rumen epithelium from birth to day 90 of age. Then, a shotgun approach and bioinformatics analyses were used to investigate and compare proteomic profiles of Holstein calve rumen epithelium on day 0, 30, 60 and 90 of age. A total of 4372 proteins were identified, in which we found 852, 342, 164 and 95 differentially expressed proteins between D0 and D30, between D30 and D60, between D60 and D90, respectively. Finally, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to provide a comprehensive proteomic landscape of dairy calves rumen development at tissue level. Conclusion: To conclude, our data indicated that keratinocyte differentiation, mitochondrion formation, the establishment of urea transport and innate immune system play central roles during rumen epithelium development. Tetrahydrobiopterin (BH4) presents an important role in rumen epithelial keratinization. The biological processes of BH4 biosynthesis and molecular function of nicotinamide adenine dinucleotide phosphate binding participate in mitochondrial cristae formation. The proposed datasets provide a useful basis for future studies to better comprehend dairy calves rumen epithelial development.

The GWAS Analysis of Body Size and Population Verification of Related SNPs in Hu Sheep.

Body size is an important indicator of growth and health in sheep. In the present study, we performed Genome-Wide Association Studies (GWAS) to detect significant single-nucleotide polymorphisms (SNPs) associated with Hu sheep's body size. After genotyping parental (G1) and offspring (G2) generation of the nucleus herd for meat production of Hu sheep and conducting GWAS on the body height, chest circumference, body length, tail length, and tail width of the two groups, 5 SNPs associated with body height and 4 SNPs correlated with chest circumference were identified at the chromosomal significance level. No SNPs were significantly correlated to body length, tail length, and width. Four out of the 9 SNPs were found to be located within the 4 genes. KITLG and CADM2 are considered as candidate functional genes related to body height; MCTP1 and COL4A6 are candidate functional genes related to chest circumference. The 9 SNPs found in GWAS were verified using the G3 generation of the nucleus herd for meat production. Nine products were amplified around the 9 sites, and 29 SNPs were found; 3 mutation sites, G > C mutation at 134 bp downstream of s554331, T > G mutation at 19 bp upstream of s26859.1, and A > G mutation at 81 bp downstream of s26859.1, were significantly correlated to the body height. Dual-luciferase reporter gene experiments showed that the 3 SNPs could significantly impact dual-luciferase and gene transcription activity.

The involvement of translationally controlled tumor protein during lamb rumen epithelium development.

Early weaning is usually applied to improve the reproductive efficiency of sheep in mutton production, while the development of rumen is of vital importance for sheep weaning age. Translationally controlled tumor protein (TCTP) is a highly conserved protein which participates in multiple tissue and organ development. Thus, we hypothesized that TCTP was involved in sheep rumen development. Histological analyses of sheep rumen epithelium showed that the epithelium formed tough shaped papillae without growing from birth to day 15 of age, after which it rapidly developed to functional epithelia on day 45 of age. We then found TCTP expressed in stratum basale, stratum spinosum and stratum granulosum of rumen epithelium. TCTP protein expression remained at a relative low level from day 0 to day 15 of age, it then significantly increased on day 30 (p < 0.05) and gradually decreased until day 60. Furthermore, to explore the role of TCTP in sheep rumen and its regulation, we found the ratio of Ki67 positive cell in stratum basale cells followed the similar pattern as the expression of TCTP. We also found the ratio of acetate:propionate in rumen fluid decreased from day 30 to day 60 of age (p < 0.05). To conclude, our data indicated that TCTP participated in rumen papillae growth by promoting rumen stratum basale cell proliferation.

Genome-Wide Association Study of Body Weights in Hu Sheep and Population Verification of Related Single-Nucleotide Polymorphisms.

Body weight (BW) is a critical economic trait for meat production in sheep. The current study aimed to perform a genome-wide association study (GWAS) to detect significant single-nucleotide polymorphisms (SNPs) that are associated with BW in Hu sheep. The comparison and analysis of the G1 and G2 generations of a nucleus meat Hu sheep breeding herd revealed four SNPs identified by GWAS. The subsequent verification of the significant SNP loci in the Hu sheep G3 generation nucleus herd also detected nine SNPs in significant SNP regions. Two SNPs were significantly associated with the BW of Hu sheep (p < 0.05). OARX_76354330.1 and s64890.1 could be identified as functional SNPs for the growth traits of Hu sheep. CAPN6, as a candidate gene, was significantly different in the biceps femoris and longissimus dorsi muscles of weaning (60-day) and 6-month sheep, which facilitated the discovery of causal variants for BW and contributed to the marker-assisted selection breeding of Hu sheep.

Enzyme Reactor Based on Reversible pH-Controlled Catalytic Polymer Porous Membrane.

The challenge for polymeric enzyme reactors at present is to selectively control the enzymolysis rate in complex conditions. Additionally, the fabrication methodology is hindered by complex processes, especially for achieving diverse stimuli responsiveness and functions. Here, we reported a kind of pH-sensitive polymer, poly(styrene- co-maleic anhydride-acrylic acid) (PS-MAn-AA)-based hybrid enzyme reactor. It comprised magnetic nanoparticles and a pH-sensitive PS-MAn-AA porous polymer membrane made by breath figure method. The enzyme l-asparaginase (l-ASNase) could covalently bond on the surface of the pH-sensitive porous polymer membrane (pH-PPM), and the resultant enzyme reactor was characterized by Fourier transform infrared spectroscopy and vibrating sample magnetometer. The apparent Michaelis-Menten constants ( Km and Vmax) of the l-ASNase enzyme reactor at different pH values were determined by a chiral ligand-exchange capillary electrophoresis method with l-asparagine as the substrate. The Vmax value of the l-ASNase enzyme reactor (0.67 mM/min) was almost 3-fold of that of the free l-ASNase (0.23 mM/min) at pH 8.2. Its ability to precisely control the enzymolysis rate in complex conditions is triggered primarily by the pH of the buffer solution, allowing controlled enzymatic reactions and displaying excellent stability and reusability of the proposed pH-PPM. This strategy for porous polymer membrane enzyme reactor fabrication has established a platform for enzyme efficiency adjusting. These valve-like distinguished features highlight the outstanding potential of stimuli-responsive enzyme reactor applied for enzyme immobilization and enzyme-related disease treatment.

A novel method to detect meat adulteration by recombinase polymerase amplification and SYBR green I.

Meat adulteration is one of the most common economic fraudulences in food industry. Current methodologies of meat source identification are complex, time-consuming and require sophisticated equipment. Hence, a simpler species specific method to determinate species is urgently needed. Here, we developed a novel method to visually identify the adulteration of meat using recombinase polymerase amplification (RPA) and SYBR green I (SG). At the isothermal temperature of 37 °C, RPA specifically identifies duck, chicken, cow, sheep and pig within 30 min of water bath. The RPA amplicons were successfully visualized by adding SG I. Furthermore, RPA can differentiate species of boiled, microwaved, high pressured or fried samples. Finally, using this system, we visually identified 1% pork adulterated in mutton or beef.

Development of alanine aminotransferase reactor based on polymer@Fe3O4 nanoparticles for enzyme inhibitors screening by chiral ligand exchange capillary electrophoresis.

Alanine aminotransferase (ALT) plays significant role in biological and clinical research. In this study, a unique ALT enzyme reactor based on multifunctional polymer@magnetic nanoparticles has been constructed for the first time and the enzymolysis efficiency has been evaluated by chiral ligand exchange capillary electrophoresis technique. Poly(N-acryloxysuccinimide) has been synthesized by reversible addition-fragmentation chain transfer polymerization method and immobilized on the magnetic nanoparticles via the succinimide group in the polymer. Interestingly, the enzyme also could easily react with the succinimide group, which enables of ALT covalent bonding onto the polymer. The enzyme amount immobilized and the immobilization time have been investigated. Comparing with free ALT in solution (Vmax of free enzyme = 0.6 mM min-1), the resultant enzyme reactor has exhibited good reusability and stability, and displayed about five times enhanced enzymolysis efficiency with L-alanine as the substrate (Vmax of enzyme reactor = 3.4 mM min-1). Furthermore, the prepared enzyme reactor has been applied in ALT inhibitors screening. The enzyme reactors based on the multifunctional polymer@magnetic nanoparticles have depicted great potential in anti-liver drugs development, liver diseases study and ALT related biological process inspect.

New chiral ligand exchange capillary electrophoresis system with chiral amino amide ionic liquids as ligands.

Using chiral amino amide ionic liquids as the ligands, a new chiral ligand exchange capillary electrophoresis method with Cu(II) as the central ion was constructed for enantioseparation of labeled D,L-amino acids. The effects of key parameters, including pH value of the running buffer, the ratio of Cu(II) to chiral amino amide ionic liquids, the concentration of complexes based on Cu(II)-chiral amino amide ionic liquids were investigated. It has been observed that eight pairs of labeled D,L-amino acids could be baseline-separated with a running buffer of 15.0mM ammonium acetate, 10.0mM Cu(II) and 20.0mML-phenylalaninamide based ionic liquid at pH 5.0. The quantitation of D,L-amino acids was conducted and good linearity (r2 ≥ 0.964) was obtained. Furthermore, an assay for determining the enantiomeric purity of D,L-amino acids was developed and the possible enantiorecognition mechanism was discussed briefly. The results indicated that the chiral amino amide ionic liquids could play the role of ligands in chiral ligand exchange capillary electrophoresis system and exhibit great potential in chiral analysis.

Weight Management Belief is the Leading Influential Factor of Weight Monitoring Compliance in Congestive Heart Failure Patients.

BACKGROUND: Daily weight monitoring is frequently recommended as a part of heart failure self-management to prevent exacerbations. This study is to identify factors that influence weight monitoring compliance of congestive heart failure patients at baseline and after a 1-year weight management (WM) program. METHODS: This was a secondary analysis of an investigative study and a randomized controlled study. A general information questionnaire assessed patient demographics and clinical variables such as medicine use and diagnoses, and the weight management scale evaluated their WM abilities. Good and poor compliance based on abnormal weight gain from the European Society of Cardiology (> 2 kg in 3 days) were compared, and hierarchical multiple logistic regression analysis was used to identify factors influencing weight monitoring compliance. RESULTS: A total of 316 patients were enrolled at baseline, and 66 patients were enrolled after the 1-year WM program. Of them, 12.66% and 60.61% had good weight monitoring compliance at baseline and after 1 year of WM, respectively. A high WM-related belief score indicated good weight monitoring compliance at both time points [odds ratio (OR), 1.043, 95% confidence interval (CI), 1.023-1.063, p < 0.001; and OR, 2.054, 95% CI, 1.209-3.487, p < 0.001, respectively). Patients with a high WM-related practice score had good weight monitoring compliance at baseline (OR, 1.046, 95% CI, 1.027-1.065, p < 0.001), and patients who had not monitored abnormal weight had poor weight monitoring compliance after the 1-year WM program (OR, 0.244, 95% CI, 0.006-0.991, p = 0.049). CONCLUSIONS: Data from this study suggested that belief related to WM plays an important role in weight monitoring compliance.

The Stimulatory Effect of Cerebral Intraventricular Injection of cNPY on Precocial Feeding Behavior in Neonatal Chicks (Gallus domesticus).

Neuropeptide Y (NPY) is one of the most potent stimulants of food intake in many animals. Most of the supporting evidence for the effects of NPY has been gathered in mammalian species using porcine NPY. To investigate the effects of NPY on precocial feeding initiation in chicks, we firstly used chicken NPY (cNPY) to study its role in food intake and spontaneous activities in 3-day-old male chicks. Food intake was monitored at different times after intracerebroventricular (ICV) injection of cNPY (2.5, 5.0 or 10.0 µg/10 µL) and anti-cNPY antibody (anti-cNPY) (1:9000, 1:3000 or 1:1000 in dilution). cNPY given at different doses significantly increased food intake at 30 min, 60 min, 90 min and 120 min after injection. Chicks treated with 5.0 µg/10 µL of cNPY showed a maximal 4.48 fold increase in food intake comparing to the control at 30 min. There is still more than 2 fold increase in food intake at 120 min after injection of cNPY. Food intake was significantly inhibited by a single ICV injection of anti-cNPY diluted to 1:9000 (60% inhibition), 1:3000 (92% inhibition), and 1:1000 (95% inhibition) at 30 min with 1:1000 being the maximally effective concentration. The inhibitory effects of anti-cNPY (diluted to1:9000, 1:3000, 1:1000) at 120 min post ICV injection were 22%, 42% and 46%, respectively. But ICV of anti-cNPY (1:3000 in dilution) did not block the orexigenic effect of 2.5 µg/10 µL of cNPY. ICV injection of different concentrations of cNPY increases locomotor activity in a dose-dependent manner while ICV anti-cNPY greatly decreased the distance moved by each chick compared to control groups. Taken together, our results demonstrated that cNPY has a promoting effect on chick food intake and locomotor activity, and that endogenous cNPY might play a positive role in regulating precocial feeding behavior in newly hatched chicks.

[Progress on miRNA in mammal breast milk].

As an important regulator in eukaryote, miRNAs could be in the animal body fluids, including serum, blood plasma, saliva, urine and so on. More recently, it was reported that miRNAs were also in the breast milk of human or cow, which indicates that miRNAs could probably be transferred into the body of the next generation by lactation and play their key roles. This might be the prelude of studies on the regulation function of miRNAs in generations. Here, we introduced the process of finding miRNAs in mammal milk, the format of miRNAs in milk and the method for isolating miRNAs, and reviewed the main functions of several miRNAs in milk. We also discussed the research task and challenge associated with miRNAs in milk at the next.

Type-dependent differential expression of neuropeptide Y in chicken hypothalamus (Gallus domesticus).

Neuropeptide Y (NPY) is one of the most important orexigenic agents in central regulation of feeding behavior, body weight and energy homeostasis in domestic chickens. To examine differences in the hypothalamic NPY between layer-type and meat-type of chickens, which are two divergent kinds of the domestic chickens in feeding behavior and body weight, we detected mRNA levels of NPY in hypothalamic infundibular nucleus (IN), paraventricular nucleus (PVN) and lateral hypothalamic area (LHA) of these two types of chickens using one-step real time RT-PCR. The meat-type chicken had more food daily (about 1.7 folds) and greater body weights (about 1.5 folds) and brain weights than the layer-type chicken at the age of 14 d. In the meat-type of chicken, NPY mRNA levels of the IN and PVN were significantly greater than those of the LHA, and were not significantly different between the IN and PVN. However, in the layer-type of chicken, NPY mRNA levels were significantly greater in the IN than those in the LHA and PVN, and were not significantly different between the PVN and LHA. In all these hypothalamic regions, the layer-type of chicken had significantly higher NPY mRNA levels than the meat-type chicken did. These results suggest the expression of NPY in the hypothalamus has a type-dependent pattern in domestic chickens.

Optimization of process parameters for reduction of gossypol levels in cottonseed meal by Candida tropicalis ZD-3 during solid substrate fermentation.

The objective of this work is to optimize the process parameters for detoxification of gossypol in cottonseed meal (CSM) by Candida tropicalis ZD-3 during solid substrate fermentation (SSF). The maximum detoxification efficiency of gossypol was achieved by employing the substrate, which consists of 70% of CSM, 20% of corn flour and 10% of wheat bran. The optimum fermentation conditions for gossypol detoxification are incubation period of 48h, incubation temperature at 30 degrees Celsius, inoculum level 5% v/w, moisture content of solid substrate 50% and pH in nature. Adding minerals solution to CSM substrate benefit fermentation detoxification.