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Abstract Memory engrams are a subset of learning activated neurons critical for memory recall, consolidation, extinction and separation. While the transcriptional profile of engrams after learning suggests profound neural changes underlying plasticity and memory formation, little is known about how memory engrams are selected and allocated. As epigenetic factors suppress memory formation, we developed a CRISPR screening in the hippocampus to search for factors controlling engram formation. We identified histone lysine-specific demethylase 4a (Kdm4a) as a negative regulator for engram formation. Kdm4a is downregulated after neural activation and controls the volume of mossy fiber boutons. Mechanistically, Kdm4a anchors to the exonic region of Trpm7 gene loci, causing the stalling of nascent RNAs and allowing burst transcription of Trpm7 upon the dismissal of Kdm4a. Furthermore, the YTH domain containing protein 2 (Ythdc2) recruits Kdm4a to the Trpm7 gene and stabilizes nascent RNAs. Reducing the expression of Kdm4a in the hippocampus via genetic manipulation or artificial neural activation facilitated the ability of pattern separation in rodents. Our work indicates that Kdm4a is a negative regulator of engram formation and suggests a priming state to generate a separate memory.

Abstract To understand the microbial diversity and community composition within the main constructive tree species, Picea crassifolia, Betula platyphylla, and Pinus tabuliformis, in Helan Mountain and their response to changes in soil physicochemical factors, a high throughput sequencing technology was used to analyze the bacterial and fungal diversity and community structure. RDA (Redundancy Analysis) and Pearson correlation analysis were used to explore the influence of soil physicochemical factors on microbial community construction, and co-occurrence network analysis was conducted on the microbial communities. The results showed that the fungal and bacterial diversity was highest in B. platyphylla, and lowest in P. crassifolia. Additionally, the fungal/bacterial richness was greatest in the rhizosphere soils of P. tabuliformis and B. platyphylla. RDA and Pearson correlation analysis revealed that NN (nitrate nitrogen) and AP (available phosphorus) were the main determining factors of the bacterial community, while NN and SOC (soil water content) were the main determining factors of the fungal community. Pearson correlation analysis between soil physicochemical factors and the alpha diversity of the microbial communities revealed a significant positive correlation between pH and the bacterial and fungal diversity, while SOC, TN (total nitrogen), AP, and AN (available nitrogen) were significantly negatively correlated with the bacterial and fungal diversity. Co-occurrence network analysis revealed that the soil bacterial communities exhibit richer network nodes, edges, greater diversity, and greater network connectivity. Indicating that bacterial communities exhibit more complex and stable interaction patterns in soil. This study reveals the complex interactive relationship between microbial communities and soil physicochemical factors in forest ecosystems. By analyzing the response of rhizosphere microbial communities of major tree species in Helan Mountain to nutrient dynamics and pH changes, we can deepen our understanding of the role of microorganisms in regulating ecosystem functions and provide theoretical basis for soil improvement and ecological restoration strategies.

IntroductionEndoglucanase (EG) and cellobiohydrolase (CBH) which produced by microorganisms, have been widely used in industrial applications.MethodsIn order to construct recombinant bacteria that produce high activity EG and CBH, in this study, eg (endoglucanase) and cbh (cellobiohydrolase) were cloned from the rumen microbial genome of yak and subsequently expressed independently and co-expressed within Lactococcus lactis NZ9000 (L. lactis NZ9000).ResultsThe recombinant strains L. lactis NZ9000/pMG36e-usp45-cbh (L. lactis-cbh), L. lactis NZ9000/pMG36e-usp45-eg (L. lactis-eg), and L. lactis NZ9000/pMG36e-usp45-eg-usp45-cbh (L. lactis-eg-cbh) were successfully constructed and demonstrated the ability to secrete EG, CBH, and EG-CBH. The sodium carboxymethyl cellulose activity of the recombinant enzyme EG was the highest, and the regenerated amorphous cellulose (RAC) was the specific substrate of the recombinant enzyme CBH, and EG-CBH. The optimum reaction temperature of the recombinant enzyme CBH was 60°C, while the recombinant enzymes EG and EG-CBH were tolerant to higher temperatures (80°C). The optimum reaction pH of EG, CBH, and EG-CBH was 6.0. Mn2+, Fe2+, Cu2+, and Co2+ could promote the activity of CBH. Similarly, Fe2+, Ba2+, and higher concentrations of Ca2+, Cu2+, and Co2+ could promote the activity of EG-CBH. The addition of engineered strains to whole-plant corn silage improved the nutritional quality of the feed, with the lowest pH, acid detergent fiber (ADF), and neutral detergent fiber (NDF) contents observed in silage from the L. lactis-eg group (p

Surface coal mining in semi-arid regions has detrimental impacts on the structure and function of surface ecosystems, thereby impeding the attainment of regional sustainable development goals. Moreover, the impact of climate change on ecological restoration in semi-arid mining areas is an inevitable consideration. To elucidate the response of ecosystem services in mining areas to regional climate change, topography, soil, vegetation and socioeconomic development, this study selected six large-scale surface coal mines located in semi-arid regions of China as research objects. In this study, we aimed to assess the main ecosystem services (carbon sequestration, soil conservation, and flow regulation) provided by these mines. In addition, we analysed the spatial and temporal evolution and interrelationships of these ecosystem services. Furthermore, we explored the underlying mechanisms between ecosystem services and environmental factors. The results showed the following: (1) Geospatially, there was a gradual decrease in carbon sequestration and flow regulation from northeast to southwest in the mining areas. However, the soil conservation exhibited an upward trend. Throughout the research period, the carbon sequestration change rate in the six mining areas displayed an initial downward and subsequent upward trend, the rate of soil conservation exhibited a gradual upward trend, and the flow regulation decline rate followed a downward trend. (2) Change rate of carbon sequestration and soil conservation in the six mining areas showed a significant positive correlation (r = 0.51, p < 0.001), with the strongest correlation observed in the Shengli Mining Area (r = 0.64, p < 0.001). (3) Ecosystem service changes in the mining areas were primarily driven by initial conditions (95.0%), followed by meteorological factors (4.2%). Three ecological restoration models were proposed for the different regions based on driver analyses. The findings of this study offer scientific evidence that can be used to inform ecological management, enhance ecological security, and promote regional sustainable development in mining areas.

Abstract Marek’s disease virus (MDV) strain GX0101 was the first reported field strain of recombinant gallid herpesvirus type 2 (GaHV-2). However, the splenic proteome of MDV-infected chickens remains unclear. In this study, a total of 28 1-day-old SPF chickens were intraperitoneally injected with chicken embryo fibroblast (CEF) containing 2000 PFU GX0101. Additionally, a control group, consisting of four one-day-old SPF chickens, received intraperitoneal equal doses of CEF. Blood and various tissue samples were collected at different intervals (7, 14, 21, 30, 45, 60, and 90 days post-infection; dpi) for histopathological, real-time PCR, and label-free quantitative analyses. The results showed that the serum expressions of MDV-related genes, meq and gB, peaked at 45 dpi. The heart, liver, and spleen were dissected at 30 and 45 dpi, and their hematoxylin-eosin staining indicated that virus infection compromised the normal organizational structure at 45 dpi. Particularly, the spleen structure was severely damaged, and the lymphocytes in the white medulla were significantly reduced. Furthermore, liquid chromatography-mass spectrometry (LC-MS) and label-free techniques were used to analyze the difference in splenic proteome profiles of the experimental and control groups at 30 and 45 dpi. Proteomic analysis identified 1660 and 1244 differentially expressed proteins (DEPs) at 30 and 40 dpi, respectively, compared with the uninfected spleen tissues. According to GO analysis, these DEPs were involved in processes such as organelle organization, cellular component biogenesis, cellular component assembly, anion binding, small molecule binding, metal ion binding, cation binding, cytosol, nuclear part, etc. Additionally, KEGG analysis indicated that the following pathways were linked to MDV-induced inflammation, apoptosis, and tumor: Wnt, Hippo, AMPK, cAMP, Notch, TGF-β, PI3K-Akt, Rap1, Ras, Calcium, NF-κB, PPAR, cGMP-PKG, Apoptosis, VEGF, mTOR, FoxO, TNF, JAK-STAT, MAPK, Prion disease, T cell receptor, and B cell receptor. We finally screened 674 DEPs that were linked to MDV infection in spleen tissue. This study improves our understanding of the MDV response mechanism in the spleen.

The legacy Global Navigation Satellite System (GNSS) satellite clock offsets obtained by the dual-frequency undifferenced (UD) ionospheric-free (IF) model absorb the code and phase time-variant hardware delays, which leads to the inconsistency of the precise satellite clock estimated by different frequencies. The dissimilarity of the satellite clock offsets generated by different frequencies is called the inter-frequency clock bias (IFCB). Estimates of the IFCB typically employ epoch-differenced (ED) geometry-free ionosphere-free (GFIF) observations from global networks. However, this method has certain theoretical flaws by ignoring the receiver time-variant biases. We proposed a new undifferenced model coupled with satellite clock offsets, and further converted the IFCB into the code and phase time-variant mixed observable-specific signal bias (OSB) to overcome the defects of the traditional model and simplify the bias correction process of multi-frequency precise point positioning (PPP). The new model not only improves the mixed OSB performance, but also avoids the negative impact of the receiver time-variant biases on the satellite mixed OSB estimation. The STD and RMS of the original OSB can be improved by 7.5–60.9% and 9.4–66.1%, and that of ED OSB (it can reflect noise levels) can be improved by 50.0–87.5% and 60.0–88.9%, respectively. Similarly, the corresponding PPP performance for using new mixed OSB is better than that of using the traditional IFCB products. Thus, the proposed pseudorange and phase time-variant mixed OSB concept and the new undifferenced model coupled with satellite clock offsets are reliable, applicable, and effective in multi-frequency PPP.

When traditional grid-forming converters directly participate in the grid-connected operation of the power grid, due to the lack of a pre-synchronization control system, the voltage amplitude and initial phase on both sides of the grid-connected point will deviate, resulting in voltage and current distortion during grid-connected mode. An active support phase-locked loop free pre-synchronization control strategy based on the third-order model of a synchronous generator is proposed to address the grid-connected problem of the grid-forming converter mentioned above. First, a model of active support control with frequency integral feedback at small signal levels was constructed. The root locus method was employed to examine how system parameters affect the stability of the active support control system. Second, by adding phase pre-synchronization controllers and amplitude pre-synchronization controllers to the active frequency loop and excitation voltage loop of the third-order model, it was ensured that the frequency, phase, and voltage amplitude of the unit are consistent with the power grid, achieving a fast and smooth grid-connected mode of the unit. Finally, by using a DC source to simulate all types of new energy power generation equipment, the active support pre-synchronization control system based on the three-order model of synchronous generator is built in the MATLAB/Simulink simulation environment, and the accuracy and effectiveness of the control strategy in this paper is verified.

The Statistical Package for Social Sciences (SPSS) Statistics software suit was used to test the model's goodness of fit and its normal distribution. Combined with the actual pavement survey data these tools were used to verify the model. The research results show that: permafrost and water-rich conditions have the same effect on pavement roughness and bumping; four key factors, including pavement riding quality index (RQI), pavement bumping index (PBI), frozen soil and water-rich environmental factors have significant impact on pavement roughness and bumping. The prediction model adjusted R2 is 0.970, which is close to 1. The proposed model provides a high degree of fit and satisfies the assumption of normal distribution. When the PBI is from 87.5 to 95, the permafrost environmental factor is from 0.0002 to 0.0014, and the water-rich environmental factor is from 10.73 to 14.87, the prediction level of the model is the best. The model's RQI prediction value and the measured value has a degree of fit 0.987, which shows a good prediction effect. The prediction model can reasonably predict the roughness and the bumping of cement concrete pavement, which is of great significance to improve road traffic safety and to prolong the service life of cement concrete pavement in seasonal frost regions and water-rich areas.

ABSTRACT: As the most prevalent pathogen of duck viral hepatitis (DVH), duck hepatitis A virus genotype 3 (DHAV-3) has caused huge economic losses to the duck industry in China. Herein, we obtained whole-transcriptome sequencing data of susceptible (S) and resistant (R) Pekin duckling samples at 0 h, 12 h, and 24 h after DHAV-3 infection. We found that DHAV-3 infection induces 5,396 differentially expressed genes (DEGs), 85 differentially expressed miRNAs (DEMs), and 727 differentially expressed lncRNAs (DELs) at 24 hpi in S vs. R ducks, those upregulated genes were enriched in inflammation and cell communications pathways and downregulated genes were related to metabolic processes. Upregulated genes showed high connectivity with the miR-33, miR-193, and miR-11591, and downregulated genes were mainly regulated by miR-2954, miR-125, and miR-146b. With the construction of lncRNA-miRNA-mRNA axis, we further identified a few aberrantly expressed lncRNAs (e.g., MSTRG.36194.1, MSTRG.50601.1, MSTRG.34328.7, and MSTRG.29445.1) that regulate expression of hub genes (e.g., THBD, CLIC2, IL8, ACOX2, GPHN, SMLR1, and HAO1) by sponging those highly connected miRNAs. Altogether, our findings defined a dual role of ncRNAs in immune and metabolic regulation during DHAV-3 infection, suggesting potential new targets for treating DHAV-3 infected ducks.

With the improvement of spatial resolution, the conveyed information of remote sensing images has become increasingly intricate. The semantic content of pixels within the same object exhibits considerable variability, whereas the semantic content of pixels between different objects exhibits significant overlap. However, most existing approaches focus solely on establishing the internal consistency of objects by aggregating global or multiscale contextual information without adequately considering the orientation and spatially detailed features of the target. Moreover, these methods often overlook the potential of edge information in achieving accurate edge positioning. These defects will adversely affect the accuracy of segmentation. In this article, we present an edge information guided network, which leverages edge information to guide the aggregation of rich contextual information for semantic segmentation to improve the segmentation accuracy of high-resolution remote sensing images. Specifically, an orientation convolution module is proposed to construct a spatial detail branch for acquiring precise edge information and spatial detail information. To effectively guide the aggregation of spatial detail features and semantic features, we propose a spatial-semantic feature aggregation module. Moreover, to enhance the extraction of long-range dependencies of irregular objects, we propose the orientation atrous convolution module, which facilitates the extraction of multiphase long-range dependencies of objects. The ISPRS Vaihingen and Potsdam datasets are employed to validate the efficacy of the proposed methodology and draw comparisons with various state-of-the-art techniques. The experimental results demonstrate that the proposed method offers distinct advantages.

Vegetation primarily draws water from soil moisture (SM), with restoration in drylands often reducing SM storage (SMS). However, anomalies have been detected in the Beijing–Tianjin Sand Source Region (BTSSR) of China via the Global Land Data Assimilation System (GLDAS) and Gravity Recovery and Climate Experiment (GRACE). This study quantified the sources of increased SMS in drylands to elucidate the effects of vegetation restoration on SMS. The results indicated the following: (1) In vegetated drylands, 46.2% experienced a significant increase in SMS while 53.8% remained stable; both were positively correlated with the normalised difference vegetation index (NDVI). (2) The increase in SMS was accompanied by a decrease in groundwater storage (GWS), as indicated by the significant correlation coefficients of −0.710 and −0.569 for SMS and GWS, respectively. Furthermore, GWS served as the primary source of water for vegetation. (3) The results of the redundancy analysis (RDA) indicated that the initial vegetation, the driver of the observed trend of increased SMS and decreased GWS, accounted for 50.3% of the variability in water storage. Therefore, to sustain dryland ecosystems, we recommend that future vegetation restoration projects give due consideration to the water balance while concurrently strengthening the dynamic monitoring of SMS and GWS.

The accurate identification of forest degradation and its driving factors is a prerequisite for implementing high-quality forest management. However, distinguishing degradation patterns is often neglected in large-scale forest quality assessments. The indicators were constructed to identify typical forest degradation patterns using remote sensing indexes, followed by an analysis of the spatiotemporal dynamics of forest degradation and quantification of the contributions from various driving factors. The results indicated that the constructed indicators could effectively distinguish typical forest degradation patterns, with a fire degradation identification accuracy of 90.0% and a fitting accuracy of drought and pest degradation higher than 0.7. The cold temperate conifer forest zone had the largest proportion of fire degradation, accounting for 67.7% of the area, and totals of 99.0% of the subtropical evergreen broadleaf forest zone and 92.8% of the temperate conifer and broadleaf mixed forest zone were moderately to severely affected by drought, with long-term stability. Additionally, 0.1% of the temperate grassland region and 0.1% of the cold temperate conifer forest zone underwent severe pest infestations, with a long-term stable trend. Meteorological factors were the primary contributors to all typical degradation patterns, accounting for 81.35%, 58.70%, and 82.29%, respectively. The research developed an index for assessing forest degradation and explained the importance of natural and anthropogenic factors in forest degradation. The results are beneficial for the scientific management of forest degradation and for improving forest management efficiency.

Matrine, an alkaloid derived from herbal medicine, has a wide range of biological activities, including antibacterial. Matrine was toxic to multiple cells at high concentrations. Bovine mammary epithelial cells (MAC-T) could be used as model cells for cow breast. Matrine was a feasible option to replace antibiotics in the prevention or treatment of mastitis against the background of prohibiting antibiotics, but the safe concentration of matrine on MAC-T cells and the mechanism of action for matrine at different concentrations were still unclear. In this study, different concentrations of matrine (0.5, 1, 1.5, 2, 2.5 and 3 mg/mL) were used to treat MAC-T cells for various time periods (4, 8, 12, 16 and 24 h) and measure their lactic dehydrogenase (LDH). And then the optimal doses (2 mg/mL) were chosen to detect the apoptosis at various time periods by flow cytometry and transcriptome analysis was performed between the control and 2 mg/mL matrine-treated MAC-T cells for 8 hours. The results showed that matrine was not cytotoxic at 0.5 mg/mL, but it was cytotoxic at 1~3 mg/mL. In addition, matrine induced apoptosis in MAC-T cells at 2 mg/mL and the proportion of apoptosis cells increases with time by flow cytometry. RNA-seq analysis identified 1645 DEGs, 676 of which were expressed up-regulated and 969 were expressed down-regulated. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated the following pathways were linked to matrine-induced toxicity and apoptosis, including cytokine-cytokine receptor interaction pathway, viral protein interaction with cytokine and cytokine receptor, P53 and PPAR pathway. We found 7 DEGs associated with matrine toxicity and apoptosis. This study would provide a basis for the safety of matrine in the prevention or treatment of mastitis.

The clustered regularly interspaced short palindromic repeat (CRISPR) is an adaptive immune system that defends most archaea and many bacteria from foreign DNA, such as phages, viruses, and plasmids. The link between the CRISPR-Cas system and the optimum growth temperature of thermophilic bacteria remains unclear. To investigate the relationship between the structural characteristics, diversity, and distribution properties of the CRISPR-Cas system and the optimum growth temperature in thermophilic bacteria, genomes of 61 species of thermophilic bacteria with complete genome sequences were downloaded from GenBank in this study. We used CRISPRFinder to extensively study CRISPR structures and CRISPR-associated genes (cas) from thermophilic bacteria. We statistically analyzed the association between the CRISPR-Cas system and the optimum growth temperature of thermophilic bacteria. The results revealed that 59 strains of 61 thermophilic bacteria had at least one CRISPR locus, accounting for 96.72% of the total. Additionally, a total of 362 CRISPR loci, 209 entirely distinct repetitive sequences, 131 cas genes, and 7744 spacer sequences were discovered. The average number of CRISPR loci and the average minimum free energy (MFE) of the RNA secondary structure of repeat sequences were positively correlated with temperature whereas the average length of CRISPR loci and the average number of spacers were negatively correlated. The temperature did not affect the average number of CRISPR loci, the average length of repeats, or the guanine-cytosine (GC) content of repeats. The average number of CRISPR loci, the average length of the repeats, and the GC content of the repeats did not reflect temperature dependence. This study may provide a new basis for the study of the thermophilic bacterial adaptation mechanisms of thermophilic bacteria.

Staphylococcus aureus (S. aureus) is one of the main pathogens causing mastitis in dairy cows. The current work mainly focuses on the pathway of apoptosis induction in MAC-T cells caused by S. aureus infection or other factors. However, the physiological characteristics of S. aureus infected MAC-T cells and the resulting mRNA expression profile remain unknown particularly in the case of diverse drug resistant strains. Methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) strains were used to infect MAC-T cells to investigate this issue. The adhesion, invasion and apoptosis ability of MRSA-infected group and MSSA-infected group was assessed over time (2, 4, 6, 8, and 12 h). After 8 h, the RNA sequencing was conducted on the MRSA-infected and the MSSA-infected with uninfected MAC-T cells as controls. The results showed that the adhesion and invasion ability of MRSA-infected and MSSA-infected to MAC-T cells increased and then decreased with infection time, peaking at 8 h. The adhesion and invasion rates of the MSSA-infected were substantially lower than those of the MRSA-infected, and the invasion rate of the MSSA-infected group was nearly non-existent. Then the apoptosis rate of MAC-T cells increased as the infection time increased. The transcriptome analysis revealed 549 differentially expressed mRNAs and 390 differentially expressed mRNAs in MRSA-infected and MSSA-infected MAC-T cells, respectively, compared to the uninfected MAC-T cells. According to GO analysis, these differentially expressed genes were involved in immune response, inflammation, apoptosis, and other processes. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated the following pathways were linked to adhesion, invasion inflammation and apoptosis, including AMPK, FOXO, HIF-1, IL-17, JAK-STAT, MAPK, mTOR, NF-κB, p53, PI3K-Akt, TNF, Toll-like receptor, Rap1, RAS, prion disease, the bacterial invasion of epithelial cells pathway. We found 86 DEGs from 41 KEGG-enriched pathways associated with adhesion, invasion, apoptosis, and inflammation, all of which were implicated in MAC-T cells resistance to MRSA and MSSA infection. This study offers helpful data toward understanding the effect of different drug-resistant S. aureus on dairy cow mammary epithelial cells and aid in the prevention of mastitis in the dairy industry.

Abstract Background People are paying more attention to the healthy and balanced diet with the improvement of their living standards. Milk fatty acids (FAs) have been reported that they were related to some atherosclerosis and coronary heart diseases in human. In our previous genome-wide association study (GWAS) on milk FAs in dairy cattle, 83 genome-wide significant single nucleotide polymorphisms (SNPs) were detected. Among them, two SNPs, ARS-BFGL-NGS-109493 and BTA-56389-no-rs associated with C18index (P = 0.0459), were located in the upstream of 1-acylglycerol-3-phosphate O-acyltransferase 3 (AGPAT3) gene. AGPAT3 is involved in glycerol-lipid, glycerol-phospholipid metabolism and phospholipase D signaling pathways. Hence, it was inferred as a candidate gene for milk FAs. The aim of this study was to further confirm the genetic effects of the AGPAT3 gene on milk FA traits in dairy cattle. Results Through re-sequencing the complete coding region, and 3000 bp of 5′ and 3′ regulatory regions of the AGPAT3 gene, a total of 17 SNPs were identified, including four in 5′ regulatory region, one in 5′ untranslated region (UTR), three in introns, one in 3′ UTR, and eight in 3′ regulatory region. By the linkage disequilibrium (LD) analysis with Haploview4.1 software, two haplotype blocks were observed that were formed by four and 12 identified SNPs, respectively. Using SAS9.2, we performed single locus-based and haplotype-based association analysis on 24 milk FAs in 1065 Chinese Holstein cows, and discovered that all the SNPs and the haplotype blocks were significantly associated with C6:0, C8:0 and C10:0 (P A, g.146704373A > G, g.146704618A > G and g.146704699G > A) changed the transcription factor binding sites (TFBSs) for transcription factors SMARCA3, REX1, VMYB, BRACH, NKX26, ZBED4, SP1, USF1, ARNT and FOXA1. Out of them, two SNPs were validated to impact transcriptional activity by performing luciferase assay that the alleles A of both SNPs, g.146704373A > G and g.146704618A > G, increased the transcriptional activities of AGPAT3 promoter compared with alleles G (P = 0.0004). Conclusions In conclusion, our findings first demonstrated the significant genetic associations of the AGPAT3 gene with milk FAs in dairy cattle, and two potential causal mutations were detected.

β-glucosidase derived from microorganisms has wide industrial applications. In order to generate genetically engineered bacteria with high-efficiency β-glucosidase, in this study two subunits (bglA and bglB) of β-glucosidase obtained from the yak rumen were expressed as independent proteins and fused proteins in lactic acid bacteria (Lactobacillus lactis NZ9000). The engineered strains L. lactis NZ9000/pMG36e-usp45-bglA, L. lactis NZ9000/pMG36e-usp45-bglB, and L. lactis NZ9000/pMG36e-usp45-bglA-usp45-bglB were successfully constructed. These bacteria showed the secretory expression of BglA, BglB, and Bgl, respectively. The molecular weights of BglA, BglB, and Bgl were about 55 kDa, 55 kDa, and 75 kDa, respectively. The enzyme activity of Bgl was significantly higher (p < 0.05) than that of BglA and BglB for substrates such as regenerated amorphous cellulose (RAC), sodium carboxymethyl cellulose (CMC-Na), desiccated cotton, microcrystalline cellulose, filter paper, and 1% salicin. Moreover, 1% salicin appeared to be the most suitable substrate for these three recombinant proteins. The optimum reaction temperatures and pH values for these three recombinant enzymes were 50 °C and 7.0, respectively. In subsequent studies using 1% salicin as the substrate, the enzymatic activities of BglA, BglB, and Bgl were found to be 2.09 U/mL, 2.36 U/mL, and 9.4 U/mL, respectively. The enzyme kinetic parameters (Vmax, Km, Kcat, and Kcat/Km) of the three recombinant strains were analyzed using 1% salicin as the substrate at 50 °C and pH 7.0, respectively. Under conditions of increased K+ and Fe2+ concentrations, the Bgl enzyme activity was significantly higher (p < 0.05) than the BglA and BglB enzyme activity. However, under conditions of increased Zn2+, Hg2+, and Tween20 concentrations, the Bgl enzyme activity was significantly lower (p < 0.05) than the BglA and BglB enzyme activity. Overall, the engineered lactic acid bacteria strains generated in this study could efficiently hydrolyze cellulose, laying the foundation for the industrial application of β-glucosidase.

Feature extraction is an essential part of automatic speech recognition (ASR) to compress raw speech data and enhance features, where conventional implementation methods based on the digital domain have encountered energy consumption and processing speed bottlenecks. Thus, we propose a Mixed-Signal Processing (MSP) architecture to efficiently extract Mel-Frequency Cepstrum Coefficients (MFCC) features. We design MSP-MFCC to pre-process speech signals in the analog domain, which significantly reduces the cost of the analog-to-digital converter (ADC), as well as the computational complexity of the digital back-end. Moreover, MSP-MFCC eliminates the time-consuming Fourier transform in the conventional digital realization by improving processing flow. We fabricated the analog part based on 180nm CMOS mixed-signal technology, then measured the chip. The measured results show the energy consumption of MSP-MFCC is $0.72~\mu \text{J}$ /frame, and the processing speed is up to $45.79~\mu \text{s}$ /frame. MSP-MFCC achieves 95% energy saving and about $6.4\times $ speedup than state of the art. Further, by using the features extracted by MSP-MFCC, speech recognition simulation reaches the accuracy of 98.2%, which also keeps the leading performance to its current counterparts. The proposed MFCC extractor is competitive for integration in the ultra-low-power always-on wearable speech recognition applications.

Abstract Background Our initial genome-wide association study (GWAS) identified 20 promising candidate genes for milk fatty acid (FA) traits in a Chinese Holstein population, including PRLR, MOGAT1, MINPP1 and CHUK genes. In this study, we performed whether they had significant genetic effects on milk FA traits in Chinese Holstein. Results We re-sequenced the entire exons and 3000 bp of the 5′ and 3′ flanking regions, and identified 11 single nucleotide polymorphisms (SNPs), containing four in PRLR, two in MOGAT1, two in MINPP1, and three in CHUK. The SNP-based association analyses showed that all the 11 SNPs were significantly associated with at least one milk FA trait (P = 0.0456 ~ 0.05). By the linkage disequilibrium (LD) analyses, we found two, one, one, and one haplotype blocks in PRLR, MOGAT1, MINPP1, and CHUK, respectively, and each haplotype block was significantly associated with at least one milk FA trait (P = 0.0456 ~ A in PRLR, and g.111599360A > G and g.111601747 T > A in MOGAT1 were predicted to alter the transcription factor binding sites (TFBSs). A missense mutation, g.39115344G > A, could change the PRLR protein structure. The g.20966385C > G of CHUK varied the binding sequences for microRNAs. Therefore, we deduced the five SNPs as the potential functional mutations. Conclusion In summary, we first detected the genetic effects of PRLR, MOGAT1, MINPP1 and CHUK genes on milk FA traits, and researched the potential functional mutations. These data provided the basis for further investigation on function validation of the four genes in Chinese Holstein.

Abstract Background We previously conducted a genome-wide association study (GWAS) strategy for milk fatty acids in Chinese Holstein, and identified 83 genome-wide significant single nucleotide polymorphisms (SNPs) and 314 suggestive significant SNPs. Among them, two SNPs, BTB-01077939 and BTA-11275-no-rs associated with C10:0, C12:0, and C14 index (P = 0.000014 ~ 0.000024), were within and close to (0.85 Mb) protein kinase, cGMP-dependent, type І (PRKG1) gene on BTA26, respectively. PRKG1 gene plays a key role in lipolysis to release fatty acids and glycerol through the hydrolysis of triacyglycerol in adipocytes. We herein considered it as a promising candidate for milk fatty acids. The purpose of this study was to investigate whether PRKG1 had effects on milk fatty acids. Results By direct sequencing the PCR products of pooled DNA, we identified a total of six SNPs, including one in 5′ flanking region, four in 3′ untranslated region (UTR), and one in 3′ flanking region. The single-locus association analysis was carried out, and showed that the six SNPs mainly had significant associations with C6:0, C8:0 and C17:1 (P A and g.6904047G > T with Haploview 4.1, and it was strongly associated with C8:0, C10:0, C16:1, C17:1, C20:0 and C16 index (P = T, was predicted to alter the binding site (BS) of transcription factor (TF) GAGA box with Genomatix software, and the subsequent luciferase assay verified that it really changed the transcriptional activity of PRKG1 gene (P = 0.0009). Conclusion In conclusion, to our best of knowledge, we are the first who identified the significant effects of PRKG1 on milk fatty acids in dairy cattle.

Placentation and placental steroidogenesis are important for pregnancy and maternal–fetal health. As pregnancy progresses, the main site of progesterone (P4) synthesis changes from the corpus luteum to the placenta, in which placental trophoblasts are the main cell type for P4 synthesis. Therefore, this study investigated the effects of P4 on apoptosis and steroidogenesis in porcine placental trophoblasts and the underlying molecular mechanisms. Porcine placental trophoblasts were treated with different concentrations of P4 for 48 h in a serum-free medium in vitro. Cell number, steroidogenesis, and relevant gene and protein expression levels were detected. A high dose of P4 (10.0 μM) significantly increased P4 (p < 0.01), androstenedione (p < 0.05), testosterone (p < 0.05), and estradiol (p < 0.05) production in porcine placental trophoblasts compared with that in control cells, while a low dose of P4 (1 × 10−3 μΜ) had no marked impact on steroid production. The mRNA expression of apoptosis-related genes (CASP3, CASP8, and Bax) (p < 0.05) and steroidogenesis-related genes (CYP11A1, CYP19A1, and StAR) (p < 0.01) was upregulated, and the expression of HSD3B and HSD17B4 was inhibited (p < 0.05) in the porcine placental trophoblasts treated with high doses of P4. Low doses of P4 had a lighter effect on gene expression than high doses. The expression of apoptosis-related proteins CASP3 (p < 0.05), and Bax (p < 0.01) and steroidogenesis-related proteins CYP19A1 (p < 0.05) and StAR (p < 0.01) was raised, but the proliferation-related protein CCND2 (p < 0.01) was downregulated in the pTr cells treated with high dose of P4. In comparison, a low dose of P4 inhibited the expression of Bax, CYP11A1 (all p < 0.01), and CCND2 (p < 0.05), but the expression of CASP3 (p < 0.05) and StAR (p < 0.01) was upregulated. In summary, excessive P4 can induce the apoptosis of porcine placental trophoblasts and lead to abnormal steroidogenesis in the placenta and hormone imbalance.

ABSTRACT: Heat stress (HS) is one of the main environmental factors affecting the efficiency of poultry production. The yellow-feather chickens (YFC) as an indigenous strain of chicken is a popular poultry breed in China. Our previous study used the RNA-seq to analyze the gene expression profiles of male YFC under HS and showed that the lipid and energy metabolism pathways are activated in livers of YFC exposed to acute HS (38°C, 4 h and 25°C recovery 2 h). In this study, we used quantitative proteome analysis based on iTRAQ to study the liver response of YFC to cycle chronic HS (38 ± 1°C, 8 h/d, 7 d, CyCHS). The male YFCs treatment used the CyCHS from 22 to 28 days of age. The liver tissue samples were collected at 28 d old. A total of 39,327 unique peptides matches were detected using iTRAQ analysis and 4,571 proteins exhibited a false discovery rate of 1% or less. Forty-six significant differentially expressed proteins (DEPs) were detected in the CyCHS group compared with the control group for the liver samples, including up- and down-regulated DEPs were 18 and 28, respectively. We found that the enriched biological process terms of the DEPs expressed in the liver were related to DNA metabolic process, oxidation-reduction process, oxidative stress and gluconeogenesis. In KEGG pathway analysis. Most of the hepatic DEPs were annotated to glutathione metabolism and TCA cycle in response to CyCHS. The up-regulation of 5 DEPs (GPX1, GSTT1, GSTT1L, RRM2, and LOC100859645) in the glutathione metabolism pathway likely reflects an attempt to deal with oxidative damage by CyCHS. The down-regulation of 3 DEPs (Isocitrate dehydrogenase [IDH3A], IDH3B, and phosphoenolpyruvate carboxykinase 1) in the TCA cycle pathway contributes to the regulation mechanism of energy metabolism and probably to cope with the balance of heat production and dissipation during CyCHS in order to adapt to high temperature environments. Our results provide insights into the potential molecular mechanism in heat-induced oxidative stress and energy in YFCs and future studies will investigate the functional genes associated with the response to HS.

Understanding the postpartum uterine involution pattern and embryonic development could facilitate bovine reproduction management, improve reproductive efficiency, and diagnosis of the reproductive disorder, which would contribute to the success of the dairy business. This study aimed to investigate postpartum uterine involution and embryonic developmental patterns or postconceptional marks of embryonic fetal development in Chinese Holstein dairy cows using B-mode ultrasonography. The results revealed a significant decline in the involution period with an increase of parity and age. The uterine involution period was shorter in multiparous cows when compared with cows with lower parities. Consistently, cows over 4 years old recovered faster than younger cows (2 or 3 years). Besides, the elder cows (over 4 years) had a relatively larger size of resumed cervix uteri and horns. Postpartum uterine involution pattern analysis revealed that the reproductive tract recovered very fast during the first 16 days postpartum for all the parity. Results of postconceptional marks of embryo development revealed a slow increase in diameter of the gravid uterine horn and crown-rump length (CRL) before day 60. In contrast, this increase was dramatic and rapid after the 60th day. We also established two models to estimate gestational age based on gravid uterine horn diameter or CRL. A formula was established to determine the gravid uterine horn size during postconceptional on day 30th–day 90th (r = 0.8714, P < 0.01). In addition, a significant positive correlation between CRL and gestational age (r = 0.98151, P < 0.01) was built. In conclusion, these results illustrated that parity and calving age had significant effects on uterine involution in Chinese Holstein cows. Crown-rump length and gravid uterine horn diameter are both efficient for evaluating the embryo growth. These current findings broaden the understanding of basic reproductive pattern in Chinese Holstein cows and could benefit bovine reproductive management primarily in postpartum and early pregnant cows to reduce the calving interval and avoid periparturient metabolic diseases.