Osteocalcin is a major non-collagenous component of the bone extracellular matrix and is considered to be an indicative factor of osteoblast differentiation. In the present study, we detected osteocalcin expression in different antler areas and growth phases by immunohisto- chemistry. Osteocalcin was highly expressed in all areas during the mineralization period and in mesenchymal cell and chondrocyte areas during the rapid growth period. The nucleotide sequence of the osteocalcin gene in sika deer antler was determined. The open reading frame was 303 bp encoding a protein of 100 amino acids. The estimated molecular mass of osteocalcin was 10.38 kDa and the theoretical isoelectric point was 5.37. The osteocalcin gene with a 6× His-tag at the C-terminus was cloned into the pGEX-4T1 vector and expressed in Escherichia coli under optimal conditions. The recombinant soluble protein fused with GST was purified with Ni-NTA resin. The purified osteocalcin protein exhibited a significant increase in HA adhesion and promoted antler chondrocyte proliferation. Osteocalcin is an important factor in regulating the rapid growth and differentiation of deer antlers.
In the current study, twenty lambs, aged 4 months, half male and half female, were classified into four groups, with five in each group. The experimental three groups of lambs were given intravenous (IV), intramuscular (IM) and subcutaneous (SC) administrations of recombinant ovine interferon-τ (roIFN-τ). The fourth group (normal control) of lambs was given normal saline injections in the same way. After administrations, blood samples were collected from the tested animals at different time points post injection, and the serum titers of roIFN-τ were measured using cytopathic effect (CPE) inhibition bioassay. The results of calculating pharmacokinetic (PK) parameters using DAS software showed that the PK characteristics of roIFN-τ through IV injection conformed to the two-compartment open model, whose half-life of distribution phases (T1/2α) was 0.33±0.034 h and the elimination half-life(T1/2β) was 5.01±0.24 h. However, the PK features of IM injection and SC injection of roIFN-τ conformed to the one compartment open model, whose Tmax were 3.11±0.26 h and 4.83±0.43 h, respectively, together with an elimination half life(T1/2β) of 9.11±0.76 h and 7. 43±0.58 h, and an absorption half-life (T1/2k(a)) of 1.13±0.31 h and 1.85±0.40 h, respectively. The bioavailability of roIFN-τ after IM administration reaches 73.57%, which is greater than that of SC administration (53.43%). These results indicate that the drug administration effect can be preferably obtained following a single dose IM administration of the roIFN-τ aqueous preparation. This study will facilitate the clinical application of roIFN-τ as a potential antiviral agent in future work.
The welfare and healthy growth of poultry under intensive feeding conditions are closely related to their living environment. In spring, the air quality considerably decreases due to reduced ventilation and aeration in cage systems, which influences the meat quality and health of broilers during normal growth stages. In this study, we analyzed the airborne bacterial communities in PM2.5 and PM10 in cage broiler houses at different broiler growth stages under intensive rearing conditions based on the high-throughput 16S rDNA sequencing technique. Our results revealed that PM2.5, PM10 and airborne microbes gradually increased during the broiler growth cycle in poultry houses. Some potential or opportunistic pathogens, including Acinetobacter, Pseudomonas, Enterococcus, Microbacterium, etc., were found in the broiler houses at different growth stages. Our study evaluated variations in the microbial communities in PM2.5 and PM10 and potential opportunistic pathogens during the growth cycle of broilers in poultry houses in the spring. Our findings may provide a basis for developing technologies for air quality control in caged poultry houses.
The Gaussian mixture model (GMM) method is popular and efficient for voice conversion (VC), but it is often subject to overfitting. In this paper, the principal component regression (PCR) method is adopted for the spectral mapping between source speech and target speech, and the numbers of principal components are adjusted properly to prevent the overfitting. Then, in order to better model the nonlinear relationships between the source speech and target speech, the kernel principal component regression (KPCR) method is also proposed. Moreover, a KPCR combined with GMM method is further proposed to improve the accuracy of conversion. In addition, the discontinuity and oversmoothing problems of the traditional GMM method are also addressed. On the one hand, in order to solve the discontinuity problem, the adaptive median filter is adopted to smooth the posterior probabilities. On the other hand, the two mixture components with higher posterior probabilities for each frame are chosen for VC to reduce the oversmoothing problem. Finally, the objective and subjective experiments are carried out, and the results demonstrate that the proposed approach shows greatly better performance than the GMM method. In the objective tests, the proposed method shows lower cepstral distances and higher identification rates than the GMM method. While in the subjective tests, the proposed method obtains higher scores of preference and perceptual quality.
To reduce the influence of the static unbalance on an infrared missile guidance system, a new static unbalance measure system for the gimbals axes has been developed. Considering the coupling effects caused by a mass eccentricity, the static balance condition and measure sequence for each gimbal axis are obtained. A novel static unbalance test approach is proposed after analyzing the dynamic model of the measured gimbal axis. This approach is to drive the measured gimbal axis to do sinusoidal reciprocating motion in a small angle and collect its drive currents in real time. Then the static unbalance of the measured gimbal axis can be obtained by the current multi-cycle integration. Also a measuring system using the proposed approach has been developed. A balanced simulator is used to verify the proposed approach by the load and repeatability tests. The results show the proposed approach enhances the efficiency of the static unbalance measurement, and the developed measuring system is able to achieve a high precision with a greater stability.
Lower Carboniferous limestone has been extracted in the “Czatkowice” open-pit hill-slope quarry in southern Poland since 1947, for the needs of metallurgical and building industries, as well as farming. We can distinguish two aquifers in the Czatkowice area: the Quaternary porous aquifer and the Carboniferous fissure-porous one. Two vertical zones representing different hydrodynamic characteristics can be indentified in the Carboniferous formations. One is a weathering zone and the other one the zone of fissures and interbedding planes. Groundwater inflows into the quarry workings have been observed at the lowest mining level (+315 m above the sea level (asl)) for over 30 years. This study concerns two hypotheses of the sources of such inflows originating either from (a) the aeration zone or from (b) the saturation zone. Inflows into the quarry combine into one stream flowing gravitationally to the doline under the pile in the western part of the quarry. This situation does not cause a dewatering need. Extending eastward mining and lowering of the exploitation level lead to increased inflows.
The technology for gob-side entry retaining in steep coal seams is still in the development stage. The analysis results of the caving structure of main roof, low influence of gateway’s stability because of long filling distance and weak dynamic effect of the gateway, and the low stress redistribution environment indicate that using this technology in steep coal seams has significant advantages. Moreover, to reinforce the waste rock and the soft floor and to better guard against the impact of the waste rock during natural filling, a rock blocking device and grouting reinforcement method were invented, and theoretical calculations result show that the blocking device has high safety factor. In addition, we also developed a set of hydraulic support devices for use in the strengthening support zone. Furthermore, because the retaining gateway was a systematic project, the selection of the size and shape of the gateway cross section and its support method during the initial driving stage is a key step. Thus, first, a section the size of bottom width and roof height of a new gateway was determined to meet any related requirements. Then, according to the cross sections of 75 statistical gateways and the support technique, it chosen a trapezoidal cross section when the dip of the coal seam is 35° < α ≤ 45°, a special and an inclined arch cross section when 45° < α ≤ 55°. Eventually, a support system of bolts and cables combined with steel mesh and steel belts was provided. The support system used optimized material and improved parameters, can enhanced the self-bearing ability of the surrounding coal and rock masses.
To keep genetic diversity, flowering plants have developed a self-incompatibility system, which can prevent self-pollination. It has been reported that calcium concentration in pistil papilla cells was increased after self-pollination in transformed self-incompatible Arabidopsis thaliana. In this study, we found that CML27 changed its expression level for both mRNA and protein when compared to transcriptome and proteome. At the same time, CML27 was expressed in the anther and pistil at a high level and reached up to 5-fold up-regulated expression in the pistil at 1 h post-pollination when compared to 0 min. In order to find out potential proteins that may interact with BoCML27, BoCML27 was expressed in and isolated from E. coli. After its co-incubation with Brassica oleracea pistil proteins, the products were separated on SDS-PAGE gels. We found a specific band at the position between 130–180 kDa. Through LC-MS-MS (Q-TOF) analysis, eight proteins were identified from the band. The proteins include 26S proteasome non-ATPase regulatory (26S), Phospholipase D, alpha 2 (PLDα2) involved in Ca2+ binding and Coatomer subunit alpha-2-like (Coatomer) involved in vesicle mediated transport. All of these identified proteins provide new insights for the self-incompatibility response in B. oleracea, specific for increasing Ca2+ concentration in pistil papilla cells.
The data aggregation process of wireless sensor networks faces serious security problems. In order to defend the internal attacks launched by captured nodes and ensure the reliability of data aggregation, a secure data aggregation mechanism based on constrained supervision is proposed for wireless sensor network, which uses the advanced LEACH clustering method to select cluster heads. Then the cluster heads supervise the behaviors of cluster members and evaluate the trust values of nodes according to the communication behavior, data quality and residual energy. Then the node with the highest trust value is selected as the supervisor node to audit the cluster head and reject nodes with low trust values. Results show that the proposed mechanism can effectively identify the unreliable nodes, guarantee the system security and prolong the network lifetime.
A novel VC (voice conversion) method based on hybrid SVR (support vector regression) and GMM (Gaussian mixture model) is presented in the paper, the mapping abilities of SVR and GMM are exploited to map the spectral features of the source speaker to those of target ones. A new strategy of F0 transformation is also presented, the F0s are modeled with spectral features in a joint GMM and predicted from the converted spectral features using the SVR method. Subjective and objective tests are carried out to evaluate the VC performance; experimental results show that the converted speech using the proposed method can obtain a better quality than that using the state-of-the-art GMM method. Meanwhile, a VC method based on non-parallel data is also proposed, the speaker-specific information is investigated using the SVR method and preliminary subjective experiments demonstrate that the proposed method is feasible when a parallel corpus is not available.
Passive source localization in shallow water has always been an important and challenging problem. Implementing scientific research, surveying, and monitoring using a short, less than ten meter long, horizontal linear array has received considerable attention in the recent years. The short array can be conveniently placed on autonomous underwater vehicles and deployed for adaptive spatial sampling. However, it is usually difficult to obtain a sufficient spatial gain for localizing long-range sources due to its limited physical size. To address this problem, a localization approach is proposed which is based on matched-field processing of the likelihood of the passive source localization in shallow water, as well as inter-position processing for the improved localization performance and the enhanced stability of the estimation process. The ability of the proposed approach is examined through the two-dimensional synthetic test cases which involves ocean environmental mismatch and position errors of the short array. The presented results illustrate the localization performance for various source locations at different signal- to-noise ratios and demonstrate the build up over time of the positional parameters of the estimated source as the short array moves at a low speed along a straight line at a certain depth.
Speaker‘s emotional states are recognized from speech signal with Additive white Gaussian noise (AWGN). The influence of white noise on a typical emotion recogniztion system is studied. The emotion classifier is implemented with Gaussian mixture model (GMM). A Chinese speech emotion database is used for training and testing, which includes nine emotion classes (e.g. happiness, sadness, anger, surprise, fear, anxiety, hesitation, confidence and neutral state). Two speech enhancement algorithms are introduced for improved emotion classification. In the experiments, the Gaussian mixture model is trained on the clean speech data, while tested under AWGN with various signal to noise ratios (SNRs). The emotion class model and the dimension space model are both adopted for the evaluation of the emotion recognition system. Regarding the emotion class model, the nine emotion classes are classified. Considering the dimension space model, the arousal dimension and the valence dimension are classified into positive regions or negative regions. The experimental results show that the speech enhancement algorithms constantly improve the performance of our emotion recognition system under various SNRs, and the positive emotions are more likely to be miss-classified as negative emotions under white noise environment.
Sapelovirus A (SV-A) is a positive-sense single-stranded RNA virus which is associated with acute diarrhea, pneumonia and reproductive disorders. The virus capsid is composed of four proteins, and the functions of the structural proteins are unclear. In this study, we expressed SV-A structural protein VP1 and studied its antigenicity and immunogenicity. SDS-PAGE analysis revealed that the target gene was expressed at high levels at 0.6 mM concentration of IPTG for 24 h. The mouse polyclonal antibody against SV-A VP1 protein was produced and reached a high antiserum titer (1: 2,048,000). Immunized mice sera with the recombinant SV-A VP1 protein showed specific recognition of purified VP1 protein by western blot assay and could recognize native SV-A VP1 protein in PK-15 cells infected with SV-A by indirect immunofluorescence assay. The successfully purified recombinant protein was able to preserve its antigenic determinants and the generated mouse anti-SV-A VP1 antibodies could recognize native SV-A, which may have the potential to be used to detect SV-A infection in pigs.
In order to compare the pathogenicity of different Tembusu virus (TMUV) strains from geese, ducks and chickens, 56 5-day-old Cherry Valley ducklings which were divided into 7 groups and infected intramuscularly with 7´105 PFU/ml per duck of six challenge virus stocks. The clinical signs, weight gain, mortality, macroscopic and microscopic lesions, virus loads in sera of 1, 3, 5, 7, 11 and 14 dpi and serum antibody titers were examined. The results showed that these viruses could make the young ducks sick, but the clinical signs differed with the different species-original strains. All the experimental groups lose markedly in weight gain compared to the control, but there were no obvious distinctions in weight gains, as well as macroscopic and microscopic lesions of dead ducks between the infected groups. However, the groups of waterfowl-derived strains (from geese and ducks) showed more serious clinical signs and higher relative expressions of virus loads in sera than those from chicken-derived. The mortality of waterfowl groups was 37.5%, and the greatest mortality of chicken groups was 12.5%. The serum antibodies of the geese-species group JS804 appeared earlier and were higher in the titers than others. Taken toghter, the pathogenicity of waterfowl-derived TMUV was more serious than chicken-derived TMUV and JS804 could be chosen as one TMUV vaccine strain to protect from the infection.