Measurement of ascorbic acid in single rat peritoneal mast cells using capillary electrophoresis with electrochemical detection

In this paper, the amount of ascorbic acid (AA) in single rat peritoneal mast cell was determined by the method of capillary electrophoresis (CE) with electrochemical detection (ED) at a carbon fiber microdisk bundle electrode. The CE–ED system and the single-cell injection system were rearranged to make the operation more convenient and efficient. In the experiment, a self-made holder made of foam was used to keep the capillary from swing, which kept the stability of the baseline of the electropherogram. The single cell was lysed completely within 5 s using the 0.1% sodium dodecylsulfate (SDS) as the cell lysis solution together with the lysis voltage of 2 kV. The quantitation analysis was accomplished by the use of calibration curves, and the amount of AA in single rat peritoneal mast cell was from 2.4 to 7.1 fmol.     

Effects of two species of inorganic arsenic on the nutrient physiology of rice seedlings

This paper reports on a hydroponics experiment that was conducted to investigate the effect of inorganic arsenics on the seedlings of the rice cultivar Shanyou63. The seedlings were subjected to two treatments, i.e., As(III) and As(V). The results showed that the morphological traits of the seedlings were significantly altered after the arsenic treatments. Analysis of nitrogen, phosphorus, potassium, and arsenic contents of the roots and leaves of the seedlings indicated that the absorption of phosphorus and potassium was mainly affected by As(III), while that of nitrogen was mainly affected by As(V). The expression of 12 genes involved in the absorption and utilization of nitrogen, phosphorus, and potassium were all observed to be down-regulated after the arsenic treatments. As(V) significantly affected the absorption and utilization of nitrogen, while As(III) significantly affected those of phosphorus and potassium. The result obtained by real-time FQ-PCR regarding the difference in the gene expressions agreed with that of our hydroponics experiment.     

Development of an α-amylase reporter system for efficient screening of clones with highly expressed heterologous protein in Hansenula polymorpha

To check feasibility and effectiveness of the α-amylase reporter system, two vectors were designed and tested using hepatitis B virus surface antigen (HBsAg) and Homo sapiens granulocyte-macrophage colony stimulating factor 2 (hGM-CSF2) as a model. By integrating the vector containing two independent cassettes into the same genome locus, high-producing clones of HBsAg (or hGM-CSF2) were screened using the α-amylase as a reporter. Results show there was a positive correlation (Correlation coefficient, R ² > 0.95) between the yield of recombinant proteins and the α-amylase activity of corresponding transformants, which was independent of the gene dosage.     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆(Glycine max)初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200 μmol·L^–1 的CuSO₄溶液进入根中, 然后在重力作用下从根基部灌注400 μmol·L^–1的K₄[Fe(CN)₆]溶液, 两种物质在根内相遇即可产生棕色的Cu₂[Fe(CN)₆]沉淀, 根据沉淀的位置来确定铜离子所经过的途径。结果表明: Cu²⁺可以穿过内皮层凯氏带, 在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀, 侧根发生的部位也产生了大量的沉淀; 当抽取K₄[Fe(CN)₆]溶液后再灌注CuSO₄溶液, 发现Cu²⁺仍然可以穿过凯氏带, 并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

Effects of partially replacing barley or corn with raw and micronised CDC SO-I oats on productive performance of lactating dairy cows

Recently, a new genotype of oat (cv. CDC SO-I, containing low-hull lignin and high-fat groat), has been developed. The objective of this study was to determine the effects of partially replacing barley and corn with the new oat and its micronisation on lactating performance of dairy cows. In a double 4 x 4 Latin square design, eight lactating dairy cows (732 +/- 46 kg body weight [BW]; parity 4 +/- 2) received total mixed rations with a forage-to-concentrate ratio of 50:50 (DM basis). The four treatments were: T1, barley only (control); T2, raw oat, replacing 42% barley of T1; T3, micronised oat, replacing 42% barley of T1; and T4, raw oat and corn blend, replacing 100% barley of T1. The results showed that dairy cows fed the new oats (T2, T3) produced more fat (p < 0.05) and more fat corrected milk (p < 0.10) than cows fed barley only (T1). The performance of cows fed the new oat and corn blend (T4) was not significantly different from other treatments. The micronisation significantly reduced protein degradability (74 vs. 63%,p < 0.05), but increased starch degradability (87 vs. 93%,p < 0.05) of the new oat. However, the overall results suggested that micronisation did not show a significant impact on milk production. The newly developed CDC SO-I oat can replace 42% barley (in T1) as a concentrate supplement in dairy total mixed rations with an increased yield of milk fat and fat corrected milk.     

Prediction of tyrosine sulfation with mRMR feature selection and analysis

Protein tyrosine sulfation is a ubiquitous post-translational modification (PTM) of secreted and transmembrane proteins that pass through the Golgi apparatus. In this study, we developed a new method for protein tyrosine sulfation prediction based on a nearest neighbor algorithm with the maximum relevance minimum redundancy (mRMR) method followed by incremental feature selection (IFS). We incorporated features of sequence conservation, residual disorder, and amino acid factor, 229 features in total, to predict tyrosine sulfation sites. From these 229 features, 145 features were selected and deemed as the optimized features for the prediction. The prediction model achieved a prediction accuracy of 90.01% using the optimal 145-feature set. Feature analysis showed that conservation, disorder, and physicochemical/biochemical properties of amino acids all contributed to the sulfation process. Site-specific feature analysis showed that the features derived from its surrounding sites contributed profoundly to sulfation site determination in addition to features derived from the sulfation site itself. The detailed feature analysis in this paper might help understand more of the sulfation mechanism and guide the related experimental validation.     

Identification of cyst wall proteins of the hypotrich ciliate <Emphasis Type="Italic">Euplotes encysticus</Emphasis> using a proteomics approach

Euplotes encysticus is a species of Hypotrich ciliates, which form cyst wall by secreting the special substances on encounter of adverse environment. It has critical significance to study the component and mechanism underlying resting cyst, during resisting unfavorable conditions in dormancy induction. The present study was aimed to investigate the effects of cyst wall proteins of Euplotes encysticus by using biochemical methods. Therefore, protein extracts were separated by SDSPAGE, identified and analyzed by MALDI-TOF MS and Bioinformatics tools. We detected 42 cyst wall proteins, 26 were functional proteins and 16 proteins consist of unknown function; which is consistent with cyst wall specificity. These results partially revealed the components of resting cyst wall formed after the cells differentiation of Euplotes encysticus. In addition, our data suggested that the function of cyst wall proteins are more likely involved in the mechanical protection, signal transduction, material transport, protein degradation and energy metabolism to survival, with potentially importance implications in the molecular mechanism of eukaryocyte dormancy under stress condition.     

Dermal mesenchymal stem cells promoted adhesion and migration of endothelial cells by integrin in psoriasis

The unusual dilatation of dermal capillaries and angiogenesis played important roles in psoriasis. Some genes and proteins of dermal mesenchymal stem cells (DMSCs) from psoriasis are abnormal and related to the function of endothelial cells (ECs). The present study was aimed to evaluate whether psoriatic DMSCs could affect adhesion and migration of ECs through neovascularization-related integrins in psoriasis. Human DMSCs, collected from psoriasis lesions and healthy skin, respectively, were cocultured with human umbilical vein endothelial cells (HUVECs). The expression levels of three integrins, that is, αvβ3, αvβ5, and α5β1 in HUVECs were tested by quantitative real-time polymerase chain reaction and Western blot analysis. The adhesion and migration of HUVECs were detected by adhesion assay and migration assay. The results showed that in psoriasis group, the expression of αVβ3 and α5β1 of HUVECs markedly increased 2.50- and 3.71-fold in messenger RNA levels, and significantly increased 1.63- and 1.92-fold in protein levels, comparing to healthy control group (all p < .05). But β5 was not significantly different between the two groups (p > .05). In addition, compared with control, psoriatic DMSCs promoted HUVECs adhesion by 1.62-fold and migration by 2.91-fold (all p < .05). In conclusion, psoriatic DMSCs impact HUVECs adhesion and migration by upregulating the expression of integrins αVβ3 and α5β1.     

Genome-wide association study identifies 7q11.22 and 7q36.3 associated with noise-induced hearing loss among Chinese population

Noise-induced hearing loss (NIHL) seriously affects the life quality of humans and causes huge economic losses to society. To identify novel genetic loci involved in NIHL, we conducted a genome-wide association study (GWAS) for this symptom in Chinese populations. GWAS scan was performed in 89 NIHL subjects (cases) and 209 subjects with normal hearing who have been exposed to a similar noise environment (controls), followed by a replication study consisting of 53 cases and 360 controls. We identified that four candidate pathways were nominally significantly associated with NIHL, including the Erbb, Wnt, hedgehog and intraflagellar transport pathways. In addition, two novel index single-nucleotide polymorphisms, rs35075890 in the intron of AUTS2 gene at 7q11.22 (combined P = 1.3 × 10⁻⁶) and rs10081191 in the intron of PTPRN2 gene at 7q36.3 (combined P = 2.1 × 10⁻⁶), were significantly associated with NIHL. Furthermore, the expression quantitative trait loci analyses revealed that in brain tissues, the genotypes of rs35075890 are significantly associated with the expression levels of AUTS2, and the genotypes of rs10081191 are significantly associated with the expressions of PTPRN2 and WDR60. In conclusion, our findings highlight two novel loci at 7q11.22 and 7q36.3 conferring susceptibility to NIHL.