Molecular characteristics of MC1R gene in tile-grey plumage of domestic chicken.

1. Tile-grey plumage is a unique and rare feather type of local chicken breeds in China, but its genetic mechanism and corresponding genes remain unknown. 2. In order to identify the genetic basis and molecular characteristics of tile-grey plumage, this experiment investigated variations of melanocortin 1 receptor (MC1R) gene in Yunnan Piao chickens with typical tile-grey plumage characteristics in contrast with three Yunnan local breeds as well as two standard breeds with different plumage colour, and analysed the association between genic variation and tile-grey plumage. 3. Through sequencing and comparison of the entire coding region of the MC1R gene, a total of 10 SNP loci were detected, of which eight were non-synonymous mutations that cause amino acid changes. The gene frequency and genotype frequency of the MC1R mutation sites in different breeds and different plumage colour groups revealed that C69T, T212C and A274G were significantly associated with tile-grey plumage. Eighteen haplotypes of the MC1R gene were constructed based on 10 nucleotide variations and eight amino acid variations. Haplotype distribution and the median joining network in breeds and plumage colour groups revealed a main haplotype (hap2) for tile-grey plumage. Hap2 is unique to the tile-grey feather of Piao chicken, and the individuals carrying this haplotype account for 62.96% of the whole tile-grey chicken. 4. The results of this study are of significance for further analysis of the molecular basis of tile-grey plumage and the selective breeding of tile-grey plumage.

[Evaluation of quality of life in patients with Graves orbitopathy and its influencing factors].

Objective: To evaluate life quality of Graves orbitopathy (GO) patients using Graves orbitopathy quality-of-life questionnaire (GO-QOL) and explore the influence factors of the quality of life of GO patients. Methods: This was a cross-sectional study conducted at The Third Affiliated Hospital of Southern Medical University including 145 newly diagnosed GO patients. All the patient answered the GO-QOL and underwent ophthalmic and endocrine assessments. The main outcome measures were the scores on GO-QOL 2 subscales: visual functioning and appearance. Based on the classification in the guideline of European Group on Graves Orbitopathy (EUGOGO), the patients were divided into two groups: mild and moderate to severe groups. Then the scales between these two groups were compared and influencing factors were analyzed. Finally, the floor and ceiling effects were assessed. Results: The GO-QOL scores for the subscales of visual functioning and appearance were 70.91±27.83 and 61.29±26.37 respectively in 145 GO patients. Visual functioning and appearance were lower in moderate to severe group (62.71±28.77 and 57.52±26.49, respectively) than in mild group (85.58±18.77 and 68.02±24.99, respectively).The GO-QOL scores for the visual functioning subscale were significantly correlated with age (P=0.002), clinical active score (P=0.011) and the degree of diplopia (P=0.00, R(2)=0.373). The GO-QOL scores for the appearance were significantly correlated with sex (P=0.05) and thyroid-stimulating hormone levels (P=0.001, R(2)=0.231). No significant ceiling or floor effects were observed for either subscale of the GO-QOL. Conclusions: With the aggravation of the disease, the quality of life of GO patients is getting worse and worse. The main influencing factors of the quality of life of GO patients include age, gender, diplopia, clinical active score and thyroid-stimulating hormone levels. Close attention needs to be paid to the quality of life of GO patients.

Unusual magnetization process and magnetocaloric effect in α-CoV2O6driven by pulsed magnetic fields.

In low-dimensional Ising spin systems, an interesting observation is the presence of step magnetization at low temperatures. Here we combine both DC and pulsed magnetic fields to study the 1/3 magnetization plateau and multiple steps in the Ising spin-chain material α-CoV2O6. Magnetization in pulsed fields is quite different from that in DC fields, showing multiple steps in an intermediate range of 4.2-6 K, inverted hysteresis below 4.2 K and asymmetric magnetization in negative fields below 11 K. We demonstrate that these unusual behaviors in magnetization are caused by the spin dynamics and the anomalous magnetocaloric effect (MCE) in α-CoV2O6, i.e., abrupt changes of sample temperature in adiabatic conditions. We successfully separate the influence between the intrinsic slow spin dynamics and the quasi-extrinsic temperature change. From the MCE, we find that some irreversible behavior is originated from the slow spin dynamics. Two different slow dynamics associated with the metastable steps are observed: one is sensitive to the slow field sweep rate at the order of ∼mT s-1and weakly depends on temperature, while the other responds to the rapid field sweep rate of ∼kT s-1and dominates at lowest temperature. We also distinguish that the metastable transition atH4is the first order and crucial for the ferrimagnetic to ferromagnetic transition. This study is useful to the understanding of multistep magnetization in α-CoV2O6and sheds light on recent experimental findings of related compounds.

Expression and characterization of a recombinant Cry1Ac crystal protein fused with an insect-specific neurotoxin ω-ACTX-Hv1a in Bacillus thuringiensis.

In order to assess possible enhancement of biopesticide activity, the fusion gene of crystal protein gene cry1Ac with the insect-specific neurotoxin ω-ACTX-Hv1a gene and egfp was expressed in Bacillus thuringiensis acrystalliferous strain Cry-B under the control of the native gene expression system. The fusion recombinant Cry-B(1Ac-ACTX-EGFP) generally produced two or three small crystal-like inclusion bodies in each cell and the GFP signal could be clearly observed. A 166 kDa full-length fusion protein was identified by immunoblot analysis. Virulence of the fusion inclusions was at least fivefold higher toward larvae of Spodoptera exigua. These results demonstrated that a foreign protein could be expressed and accumulate as parasporal inclusions in B. thuringiensis by C-terminal fusion with the native endotoxin while retaining partial insecticidal activity.

Purification and characterization of the recombinant form of Acyl CoA oxidase 3 from the yeast Yarrowia lipolytica.

The Acyl CoA dependent oxidase 3 (Aox3p) from the yeast Yarrowia lipolytica, expressed in Escherichia coli, as an active protein with a 6 His tag at its N-terminal region has been purified to electrophoretic homogeneity. The purified enzyme exhibits a specific activity of 1.95 microM/min/mg using hexanoyl-CoA as substrate, and it remains active for at least 1 month upon storage at -30 degrees C in the presence of 35% (V/V) glycerol. The pH and temperature optima of the enzyme are 7.4 and 28-38 degrees C, respectively. Aox3p catalyzes the oxidation of both aliphatic acyl-CoA substrates of different chain lengths (e.g., hexanoyl-CoA, decanoyl-CoA, myristyl-CoA) as well as of the aromatic/heterocyclic ring-substituted chromogenic substrates, such as furylpropionyl-CoA. Of the above substrates, the efficiency of the enzyme, as judged by its kcat to Km ratio, exhibits the following order: decanoyl CoA > myristyl CoA > hexanoyl CoA > furyl-propionyl-CoA (FPCoA). Phenol, which is normally used in the coupled assay system for monitoring the H2O2 formation, functions as both an activator (at low concentrations) and a competitive inhibitor (at high concentrations) with respect to acyl-CoA substrates. The magnitude of activation and inhibition of the enzyme is dependent on the nature of the acyl-CoA substrates.