Polymorphism Analysis of the Agouti-related Protein Gene Exon Ⅰof Quail

The poultry feather color inheritance has been a hot research topic, because the feather color is a kind of important quality character.In this study, we used polyacrylamide gel electrophoresis to detect the polymorphisms of four quail populations of different feather color (China Yellow quail, Black quail, Korean quail, Beijing White quail) of agouti-related protein gene (Agrp) exon Ⅰ, so as to investigate the relationship of Agrp exon I with quail plumage color, and to provide reference for the breeding and production of quail.The results showed that two alleles were detected in the Agrp exon Ⅰ of China Yellow quail, Black quail, Korean quail, Beijing White quail populations,and expressed by A and B, respectively. Frequency of allele A in Beijing White quail was up to 0.9500. The frequency of B allele in Korean quail was the highest, reaching 0.3392, in Beijing White quail genes in the lowest frequency is only 0.0500. The genotype frequency of AA genotype in Beijing White quail was the highest (0.9500), the lowest in Korean quail and Black quail (0.4286). BB genotype was not observed in Beijing White quail. We speculated that it may have a certain relationship between Agrp exon Ⅰ B allele and feather color of quail.     

Research on the Effect of Plant Combination Types on the Shear Strength of the Root-soil Composite System of Alpine Grassland in the Source Region of the Yellow River北大核心CSCD

In this study,an alpine grassland in Henan County,Qinghai Province,in the source region of the Yellow River,was selected as the study area. The shear strength of root-soil composite system characteristics of 6 different plant combination types of grassland were explored by field investigation and sampling and indoor tests,which aimed to provide a theoretical basis for preventing and controlling soil erosion and shallow landslide caused by grassland degradation. The test results showed that the shear strength of root-soil composite system of 6 different combinations was a increasing trend with the decrease of altitude. Namely,the shear strength of the root-soil composite system of Stipa purpurea Griseb. + Kobresia pygmaea C. B. Clarke at the top of the sunny slope was 27.76 kPa and the value of Elymus nutans Griseb. at the bottom of the sunny slope was 37.53 kPa. The reasons why the shear strength of 6 kinds of combinations varied from large to small with the decrease of altitude were related to their density,moisture content,root content and dry weight of roots in the root-soil composite system samples. That is,the root content of composite system at the top,middle and bottom of the sunny slope were 6.2%,5.5% and 16.2%,respectively. The values at the top,middle and bottom of the shady slope were 8.0%,11.4% and 31.6%,respectively. © 2019 China Agricultural University. All rights reserved.     

Mining the key regulatory genes of chicken inosine 5′-monophosphate metabolism based on time series microarray data

IMP (inosine 5′-monophosphate) is a compound that enhances the flavor of poultry meat. IMP has become a new breeding trait to improve poultry meat quality. We tried to identify several potential regulatory genes, and construct their predicted regulatory relationships. Time series gene expression profiles of thigh muscle tissues of Rugao chicken, a famous indigenous breed in China, were performed for analysis of genes that are co-expressed or correlated with the concentration of IMP. We found 15 crucial co-expression genes, which are Hspa2, Pten, Gabpa, Bpi, Mkl1, Srf, Cd34, Hspa4, Etv6, Bmpr2, Gde1, Igfbp5, Cd28, Pecam1 and Gja1, that may directly or indirectly regulate IMP metabolism. Eventually, we computed the correlation coefficient between 19 IMP Genes and 15 CGs (15 co-expression genes), and we identified and constructed a predicted regulation network. In conclusion, variation of IMP concentration was primarily connected with the muscle development process. During this process, 15 CGs were identified that may have significant influence on IMP metabolism. In particular, Bmpr2, Pten and co-expression genes correlated with Entpd8 might play important roles in regulating IMP de novo synthesis, decomposition and salvage synthesis.

Electronic supplementary material

The online version of this article (doi:10.1186/s40104-015-0022-3) contains supplementary material, which is available to authorized users.     

Functional Role of Feline Zona Pellucida Protein 4 Trefoil Domain: A Sperm Receptor or Structural Component of the Domestic Cat Zona Pellucida?

The trefoil domain (TFD) is a protein structure characterized by six cysteines, which form a typical three-loop structure by three disulphide bridges. It is assumed that two of these loops generate a hydrophobic groove, which could be a binding site for carbohydrate residues or proteins. The zona pellucida (ZP) protein, ZP4, contains such a TFD. The carbohydrate-/protein-binding property of TFD allows us to assume a potential sperm receptor function of this domain. Additionally, gastrointestinal trefoil peptides are stable against proteases; therefore, a structural role of TFD within the ZP might also be possible. We were able to show that the synthesized and natural folded feline TFD (fTFD) expresses the typical protease resistance that vanished under reducing conditions and after substitution of cysteine residues within the peptide. Furthermore, an antibody directed against the first loop of fTFD was almost unable to bind to intact in vitro mature cat oocytes. Pre-incubation of oocytes in the reducing agent (DDT), however, improved antibody binding substantially. Therefore, we suggest structural masking of the fTFD domain within the intact ZP. An interaction between fTFD and feline sperm cells was examined using several methods, including immunocytochemistry, immunoelectron microscopy, co-immunoprecipitation and far western blot, but we found no indication for an involvement of TFD in the primary sperm binding to the ZP. To summarize, there is increasing evidence that the TFD of fZP4 has a structural rather than a sperm-binding function.     

Effect of Dietary Nonstructural Carbohydrate Content on Activation of 5′‐Adenosine Monophosphate‐Activated Protein Kinase in Liver,Skeletal Muscle,and Digital Laminae of Lean and Obese Ponies

Background

In EMS‐associated laminitis, laminar failure may occur in response to energy failure related to insulin resistance (IR) or to the effect of hyperinsulinemia on laminar tissue. 5′‐Adenosine‐monophosphate‐activated protein kinase (AMPK) is a marker of tissue energy deprivation, which may occur in IR.

Hypothesis/Objectives

To characterize tissue AMPK regulation in ponies subjected to a dietary carbohydrate (CHO) challenge.

Animals

Twenty‐two mixed‐breed ponies.

Methods

Immunohistochemistry and immunoblotting for total AMPK and phospho(P)‐AMPK and RT‐qPCR for AMPK‐responsive genes were performed on laminar, liver, and skeletal muscle samples collected after a 7‐day feeding protocol in which ponies stratified on body condition score (BCS; obese or lean) were fed either a low‐CHO diet (ESC + starch, approximately 7% DM; n = 5 obese, 5 lean) or a high‐CHO diet (ESC + starch, approximately 42% DM; n = 6 obese, 6 lean).

Results

5′‐Adenosine‐monophosphate‐activated protein kinase was immunolocalized to laminar keratinocytes, dermal constituents, and hepatocytes. A high‐CHO diet resulted in significantly decreased laminar [P‐AMPK] in lean ponies (P = .03), but no changes in skeletal muscle (lean, P = .33; obese, P = .43) or liver (lean, P = .84; obese, P = .13) [P‐AMPK]. An inverse correlation existed between [blood glucose] and laminar [P‐AMPK] in obese ponies on a high‐CHO diet.

Conclusions and Clinical Importance

Laminar tissue exhibited a normal response to a high‐CHO diet (decreased [P‐AMPK]), whereas this response was not observed in liver and skeletal muscle in both lean (skeletal muscle, P = .33; liver, P = .84) and obese (skeletal muscle, P = .43; liver, P = .13) ponies.     

A Comparison of the Concentrations of C-reactive Protein and α1-Acid Glycoprotein in the Serum of Young and Adult Dogs with Acute Inflammation

The concentrations of C-reactive protein (CRP) and ₁-acid glycoprotein (AAG) were evaluated in 1-, 3- and 18-month-old dogs (four of each age) that had been inoculated with turpentine oil. The CRP and AAG in 3-month-old and younger dogs subjected to surgery or inoculated with either Staphylococcus aureus or a viral vaccine were also evaluated. The average CRP concentration in the sera peaked 2 days after inoculation of turpentine oil. The peak CRP concentrations in 3- and 18-month-old dogs were significantly (p<0.05) greater than those in 1-month-old dogs. The average AAG concentration in the sera peaked 4 days after inoculation of turpentine oil. No significant difference was found in AAG concentrations between any of the age groups. When experimentally inoculated with S. aureus or subjected to oophorohysterectomy, the CRP and AAG concentrations increased in 3-month-old dogs, but they increased little in 1-month-old dogs. The CRP and AAG in dogs inoculated with the viral vaccine did not increase. In dogs with fractures or subjected to percutaneous gastrostomy, the CRP and AAG concentrations correlated with the condition of dogs.     

Toluene Exposure Leads to a Change in Expression Patterns of β Defensins in the Mouse Tracheal Epithelium

Defensins are generally implicated in the quick resistance of epithelial surfaces to microbials; however, recent reports have indicated that defensins also have unknown purposes in relation to noninfectious diseases. In this study, the localization patterns of anti-microbial peptides, β defensins (BDs), in the tracheal epithelium of male C3H mice under exposure to toluene were analyzed by immunohistochemistry. Mice were exposed one to ten times to toluene for 30 min by nose-only inhalation. Expression of BDs was revealed by immunohistochemistry in serial sections of trachea after the final exposure. Expression of BD-1 was usually observed at almost the same levels in all exposure groups, and expression of BD-2 was observed in the control group; however, the signals for BD-2 decreased gradually with frequency of exposure. In the group exposed ten times, expression of BD-2 decreased to far lower than that of the control group. No expression of BD-3 was detected in any groups. Interestingly, expression of BD-4 increased to the maximum in the group exposed four times and decreased to a level lower than that of the control in the group exposed ten times. The results of the present study indicated that toluene gas might change the expression pattern of BDs in the tracheal epithelial cells. The oscillation of expression of BD-4 was quite characteristic and might contribute to morphological damage in on the epithelial cells.     

Effects of Litter Addition on Soil Organic Carbon Mineralization in the Degraded Alpine Meadow in the Three-River Headwaters Region北大核心CSCD

To explore the effects of plant community structure changes on soil organic carbon mineralization in the different degraded alpine meadows in the Three-River Headwaters Region, we collected topsoil from different degraded alpine meadows, and then added three kinds of litters from different functional groups (grasses, sedges, and forbs) incubated in the laboratory for 203 days. The results showed that the cumulative mineralization of soil organic carbon first increased and then decreased as the degree of soil degradation increased. Litter addition significantly increased the cumulative mineralization of soil organic carbon of non-degraded, slightly degraded, moderately degraded, and severely degraded meadow by 102.04%, 58.71%, 70.68%, and 181.32%, respectively. However, there was no significant difference among the positive effects of different functional groups. Regression analysis showed that soil organic carbon, total nitrogen, and pH of different degraded alpine meadows could better explain the changes in the cumulative mineralization of soil organic carbon. The cumulative mineralization of soil organic carbon first increased and then decreased with soil organic carbon, total nitrogen and pH increased, while the influence degree and direction were altered by litter addition. In general, the plant community structure changes in the different degraded alpine meadows in the Three-River Headwaters Region significantly influenced soil organic carbon mineralization. © 2019 Authors. All rights reserved.     

Effect of LDL on the Mitochondrial Transmembrane Potential and ZP Protein Ubiquitination in Vitrified-warmed MⅡ Oocytes

The purpose of this study was to determine the effect of LDL addition on oocyte developmental rate,mitochondrial membrane potential and ZP protein ubiquitination level in MⅡ oocytes from vitrified-warmed GV oocytes.Mitochondrial membrane potentials were labeled with specific probe JC-1,and ZP protein samples of different treatment groups in MⅡ oocytes were analyzed by SDS-PAGE and Western blotting methods. The results showed that normal rate and in vitro maturation of MⅡ oocytes in the 10 mg/mL LDL group (71.92% and 69.86%) were significantly higher than those in control group (58.26% and 54.55%)(P<0.05),that were nearest to non-vitrified group.The mitochondrial membrane potential of MⅡ oocytes in the 10 mg/mL LDL group was significantly higher than those in control group and 1,20 mg/mL LDL groups (P<0.05).The ubiquitinated ZP1,ZP2 and ZP3 of MⅡ oocytes in non-vitrified group and LDL treated groups were labeled at 61,80 and 106 ku,respectively.The ubiquitinated ZPs (ZP1,ZP2 and ZP3) level of MⅡ oocytes in the 10 mg/mL LDL group was significantly higher than those in 1 and 20 mg/mL LDL groups (P<0.05),while significantly lower than that of non-vitrified group (P<0.05). In conlusion, LDL could improve the maturation rate, mitochondrial membrane potential and ubiquitinated ZPs level of MⅡ oocytes from vitrified-warmed GV oocytes.     

Characterization of the gut microbiota in leptin deficient obese mice – Correlation to inflammatory and diabetic parameters

Gut microbiota have been implicated as a relevant factor in the development of type 2 diabetes mellitus (T2DM), and its diversity might be a cause of variation in animal models of T2DM. In this study, we aimed to characterise the gut microbiota of a T2DM mouse model with a long term vision of being able to target the gut microbiota to reduce the number of animals used in experiments. Male B6.V-Lep^ob/J mice were characterized according to a number of characteristics related to T2DM, inflammation and gut microbiota. All findings were thereafter correlated to one another in a linear regression model. The total gut microbiota profile correlated to glycated haemoglobin, and high proportions of Prevotellaceae and Lachnospiraceae correlated to impaired or improved glucose intolerance, respectively. In addition, Akkermansia muciniphila disappeared with age as glucose intolerance worsened. A high proportion of regulatory T cells correlated to the gut microbiota and improved glucose tolerance. Furthermore, high levels of IL-10, IL-12 and TNF-α correlated to impaired glucose tolerance, blood glucose or glycated haemoglobin. The findings indicate that gut microbiota may contribute to variation in various disease read-outs in the B6.V-Lep^ob/J model and considering them in both quality assurance and data evaluation for the B6.V-Lep^ob/J model may have a reducing impact on the inter-individual variation.     

Study on the Activity Estimation Methods of Recombinant Chicken Interferon-α/Interleukin-2 Fusion Protein in vitro

In order to establish a stable,simple and rapid detection method to estimate the activities of the recombinant chicken interferon-α/interleukin-2 fusion protein (rChIFN-α-Linker-ChIL-2,recombinant fusion protein) in vitro,the activities of rChIFN-α-Linker-ChIL-2 were estimated by detecting its specific immune response to monoclonal antibody (MAb) against ChIFN-α and ChIL-2 by ELISA assay.The antiviral activities of rChIFN-α-Linker-ChIL-2 protein were tested by inhibiting the 50% appearance of cytopathic effect (CPE) of vesicular stomatitis virus (VSV) and infectious bursal disease virus (IBDV) on the passage cell lines DF1.The promoting proliferation activities of lymphocytes in the chicken peripheral blood and spleen of recombinant fusion protein were tested by MTS method.The results showed that rChIFN-α-Linker-ChIL-2 protein had the ability of specific immune response to anti-ChIFN-α MAb and anti-ChIL-2 MAb,respectively.The antiviral activity of rChIFN-α-Linker-ChIL-2 protein inhibiting the reproduction of VSV on DF1 cell line was higher than IBDV,and both of the antiviral activities of recombinant fusion protein against VSV and IBDV were much higher than the recombinant ChIFN-α protein (rChIFN-α) control.The recombinant fusion protein had apparent promoting proliferation activity of lymphocytes in the chicken peripheral blood and spleen,which were much higher than that of the rChIFN-α control.The study suggested that the activities detection and estimation methods of the recombinant fusion protein in vitro were successfully established,which laid the foundation for the further study of the synergy activity of recombinant fusion protein in vivo.     

Comparison the effects of 5-Aza-2′-deoxycytidine and zebularine on the in vitro development,blastocyst quality,methylation pattern and conception rate on handmade cloned buffalo embryos

In this study we treated the handmade cloned (HMC) buffalo embryos with the DNA methylation inhibitors; 5-aza-2′-deoxycytidine (AzadC) or Zebularine individually after post-fusion and during in vitro culture till eighth day. The blastocysts production rate significantly improved (p < .01) after treating embryos independently with 5 nM AzadC and 5 nM zebularine compared with 2 and 10 nM AzadC or zebularine groups, respectively. The highest cleavage rates were obtained for 5 nM treatment of AzadC and zebularine compared with other treatments and untreated control group. Quality of blastocysts were evaluated using total cell number (TCN) and the ratio of number of inner cell mass (ICM) cells/total cell number (ICM/TCN). Zebularine treatments (2/5/10 nM) significantly improved both TCN and ICM/TCN ratio compared with AzadC treatments (2/5/10 nM); however, control group TCN and ICM/TCN ratio was found lower. The methylation percentage of pDS4.1 and B. bubalis satellite DNA were comparatively more attenuated with 5 nM zebularine than 5 nM AzadC treatment. The increased in vitro development rates of the treated embryos were correlated with the decreased level of DNA methylation and the improved blastocyst quality. Following transfer of 5 nM zebularine treated embryos to 6 recipients, 4 were found to be pregnant, though the pregnancies were not carried to full term.     

抗肝片吸虫病药物化合物：N—（2′，4′，5′—三氯）苯基—2—羟基—3，5，6—三氯苯磺酰胺的杀虫活性研究

本文研究了治疗牛、羊等家畜肝片吸虫病药物N－（2′，4′，5′－三氯）苯基－2羟基－3，5，6－三氯－苯磺酰胺化合物的杀虫机理，通过实验及药物分子的结构分析，初步认为其杀虫机理是对线粒体氧化磷酸化反应的解偶联作用；采用氧电极法、测线粒体呼吸过程中磷的变化以及测ATP酶活性等方法研究其对线粒氧化磷酸化反应的解偶联作用，结果表明：标题药物化合物是较好的解偶联剂，其解偶联活性高于典型的解偶联剂DNP。     

The composition of the soil seedbank and its role in ecosystem dynamics and rehabilitation potential in the arid Tankwa Karoo Region,South Africa§

Very few soil seedbank studies have been conducted in South Africa, especially in arid rangelands. Insight into the soil seedbank could therefore improve assessment of rangeland dynamics and enhance rehabilitation efforts. This study aimed to characterise the soil seedbanks in various vegetation types of the Tankwa Karoo National Park, an arid environment in South Africa. At 43 sites soil was sampled twice at the end of spring and the end of autumn, and the composition and structure of aboveground vegetation were described at the peak growth period in spring. Seeds were isolated from the soil samples by means of flotation in a salt solution, seedbank species composition was obtained by seedling germination, and the vegetation was investigated using a line-point survey method. Overall seed density was 8 034 seeds m^?2 of which 55% was viable, and the common life-forms were therophytes and chamaephytes. Species richness and diversity were lower in the seedbank compared with those of the vegetation, and the two vegetation spheres were 25% similar. While seedbank composition suggested poor veld condition, there was enough seed density and viability for future regeneration and rehabilitation initiatives. However, this may be impeded by the absence of many perennial species in the soil seedbank.     

Flow Cytometric Characterization and Clinical Outcome of CD4+ T‐Cell Lymphoma in Dogs: 67 Cases

Background

Canine T‐cell lymphoma (TCL) is conventionally considered an aggressive disease, but some forms are histologically and clinically indolent. CD4 TCL is reported to be the most common subtype of TCL. We assessed flow cytometric characteristics, histologic features when available, and clinical outcomes of CD4+ TCL to determine if flow cytometry can be used to subclassify this group of lymphomas.

Objective

To test the hypothesis that canine CD4+ T‐cell lymphoma (TCL) is a homogeneous group of lymphomas with an aggressive clinical course.

Animals

Sixty‐seven dogs diagnosed with CD4+ TCL by flow cytometry and treated at 1 of 3 oncology referral clinics.

Methods

Retrospective multivariable analysis of outcome in canine CD4+ TCL including patient characteristics, treatment, and flow cytometric features.

Results

The majority of CD4+ TCL were CD45+, expressed low class II MHC, and exhibited an aggressive clinical course independent of treatment regimen (median survival, 159 days). Histologically, CD4+ TCL were classified as lymphoblastic or peripheral T cell. Size of the neoplastic lymphocytes had a modest effect on both PFI and survival in this group. A small number of CD4+ TCL were CD45− and class II MHC high, and exhibited an apparently more indolent clinical course (median survival not yet reached).

Conclusions and Clinical Importance

Although the majority of CD4+ TCL in dogs had uniform clinical and flow cytometric features and an aggressive clinical course, a subset had a unique immunophenotype that predicts significantly longer survival. This finding strengthens the utility of flow cytometry to aid in the stratification of canine lymphoma.