Combination effects of chronic cadmium exposure and gamma-irradiation on the genotoxicity and cytotoxicity of peripheral blood lymphocytes and bone marrow cells in rats

This work investigated the effects of chronic cadmium (Cd) exposure combined with γ-ray irradiation on the cytotoxicity and genotoxicity of peripheral blood cells and bone marrow cells in rats. Results showed that when the rats were exposed to low dose (LD) Cd of 0.1mg CdCl?/(kgd) for 8 and 12 weeks, the Cd concentration in blood reached to 135-140 μg/L and no toxic effects on peripheral blood lymphocytes, white blood cells (WBC) and granulocyte-monocyte (GM) progenitor cells were observed except polychromatic erythrocytes (PCE) of bone marrow. Moreover, this chronic LD Cd exposure significantly decreased irradiation-induced micronucleus (MN) formation and hypoxanthine-guanine phosphoribosyl transferase (hprt) mutation in lymphocytes and PCE, while the combination of LD Cd exposure and irradiation induced the additive metallothionein (MT) protein expression in bone marrow cells. When the rats were exposed to a high dose (HD) Cd of 0.5mg CdCl/?(kgd) for 8 and 12 weeks, the blood Cd level approached to 458-613 μg/L and an inflammatory response was induced, meanwhile, MN formation and hprt mutation were markedly increased, and the ratio of PCE/NCE (normochromatic erythrocyte) was significantly decreased. Furthermore, when the rats were exposed to HD Cd plus 2 Gy irradiation, additive toxic effects on MN formation, hprt mutation, PCE damage and GM progenitor cell proliferation were observed, while this combination treatment resulted in an obvious reduction of MT protein compared to HD Cd group. In conclusion, chronic exposure to LD Cd induced the adaptive response to irradiation in the genotoxicity of peripheral blood lymphocytes and PCE of bone marrow by the up-regulation of Cd-induced MT protein, but the combination of HD Cd exposure and irradiation generated the additive effects on the cytotoxicity and genotoxicity in peripheral blood lymphocytes and bone marrow cells.     

The First Extracellular Domain Plays an Important Role in Unitary Channel Conductance of Cx50 Gap Junction Channels

Gap junction (GJ) channels provide direct passage for ions and small molecules to be exchanged between neighbouring cells and are crucial for many physiological processes. GJ channels can be gated by transjunctional voltage (known as V_j-gating) and display a wide range of unitary channel conductance (γ_j), yet the domains responsible for V_j-gating and γ_j are not fully clear. The first extracellular domain (E1) of several connexins has been shown to line part of their GJ channel pore and play important roles in V_j-gating properties and/or ion permeation selectivity. To test roles of the E1 of Cx50 GJ channels, we generated a chimera, Cx50Cx36E1, where the E1 domain of Cx50 was replaced with that of Cx36, a connexin showing quite distinct V_j-gating and γ_j from those of Cx50. Detailed characterizations of the chimera and three point mutants in E1 revealed that, although the E1 domain is important in determining γ_j, the E1 domain of Cx36 is able to effectively function within the context of the Cx50 channel with minor changes in V_j-gating properties, indicating that sequence differences between the E1 domains in Cx36 and Cx50 cannot account for their drastic differences in V_j-gating and γ_j. Our homology models of the chimera and the E1 mutants revealed that electrostatic properties of the pore-lining residues and their contribution to the electric field in the pore are important factors for the rate of ion permeation of Cx50 and possibly other GJ channels.     

Cell type-specific proteomics uncovers a RAF15-SnRK2.6/OST1 kinase cascade in guard cells

Multicellular organisms such as plants contain various cell types with specialized functions. Analyzing the characteristics of each cell type reveals specific cell functions and enhances our understanding of organization and function at the organismal level. Guard cells (GCs) are specialized epidermal cells that regulate the movement of the stomata and gaseous exchange, and provide a model genetic system for analyzing cell fate, signaling, and function. Several proteomics analyses of GC are available, but these are limited in depth. Here we used enzymatic isolation and flow cytometry to enrich GC and mesophyll cell protoplasts and perform in-depth proteomics in these two major cell types in Arabidopsis leaves. We identified approximately 3,000 proteins not previously found in the GC proteome and more than 600 proteins that may be specific to GC. The depth of our proteomics enabled us to uncover a guard cell-specific kinase cascade whereby Raf15 and Snf1-related kinase2.6 (SnRK2.6)/OST1(open stomata 1) mediate abscisic acid (ABA)-induced stomatal closure. RAF15 directly phosphorylated SnRK2.6/OST1 at the conserved Ser175 residue in its activation loop and was sufficient to reactivate the inactive form of SnRK2.6/OST1. ABA-triggered SnRK2.6/OST1 activation and stomatal closure was impaired in raf15 mutants. We also showed enrichment of enzymes and flavone metabolism in GC, and consistent, dramatic accumulation of flavone metabolites. Our study answers the long-standing question of how ABA activates SnRK2.6/OST1 in GCs and represents a resource potentially providing further insights into the molecular basis of GC and mesophyll cell development, metabolism, structure, and function.     

The accumulation of pigment in fiber related to proanthocyanidins synthesis for brown cotton

Brown cotton is a kind of naturally colored cotton. Because of less processing and little dying, it is more friendlier to environment than white cotton. For brown cotton, pigment accumulation in fiber is one of the most important characteristics. In this study, we selected a brown fiber line and a white fiber cultivar to determine the factor that affects the pigmentation in brown fiber. Accordingly, fibers were collected to verify the presence of PAs by p-dimethylaminocinnamaldehyde (DMACA) and toluidine blue O (TBO) staining. The PAs content and related genes expressions were determined. As a result, there were obvious differences on the aspect of PAs synthesis in fiber between white cotton and brown cotton. For white fiber, the PAs content reached maximum at 5 DPA, and then gradually decreased to zero. But for brown fiber, the PAs content was increased from 5 to 15 DPA stage, and reached the maximum at the 15 DPA stage, then gradually decreased from 15 to 40 DPA stage. On the contrary, in white cotton, PAs were synthesized in the whole developmental stage from 5 to 40 DPA. And PAs content in brown fiber were far more than that in white fiber, which may be the reason why the brown pigment accumulated in brown fiber.     

Multi-representation of symbolic and nonsymbolic numerical magnitude in Chinese number processing

Numerical information can be conveyed by either symbolic or nonsymbolic representation. Some symbolic numerals can also be identified as nonsymbolic quantities defined by the number of lines (e.g., I, II, III in Roman and -, =, ≡ in Japanese Kanji and Chinese). Here we report that such multi-representation of magnitude can facilitate the processing of these numerals under certain circumstances. In a magnitude comparison task judging 1 to 9 (except 5) Chinese and Arabic numerals presented at the foveal (at the center) or parafoveal (3° left or right of the center) location, multi-representational small-value Chinese numerals showed a processing advantage over single-representational Arabic numerals and large-value Chinese numerals only in the parafoveal condition, demonstrated by lower error rates and faster reaction times. Further event-related potential (ERP) analysis showed that such a processing advantage was not reflected by traditional ERP components identified in previous studies of number processing, such as N1 or P2p. Instead, the difference was found much later in a N400 component between 300-550 msec over parietal regions, suggesting that those behavioral differences may not be due to early processing of visual identification, but later processing of subitizing or accessing mental number line when lacking attentional resources. These results suggest that there could be three stages of number processing represented separately by the N1, P2p and N400 ERP components. In addition, numerical information can be represented simultaneously by both symbolic and nonsymbolic systems, which will facilitate number processing in certain situations.     

Characteristics and metabolic pathway of Alcaligenes sp. TB for simultaneous heterotrophic nitrification-aerobic denitrification

Applied Microbiology and Biotechnology - A novel heterotrophic nitrification-aerobic denitrification bacterium, Alcaligenes sp. TB (GenBank accession no. JQ044686), was isolated from a rotating...     

猪VHL基因克隆及敲低克隆胚胎的构建

通过在细胞和胚胎水平对猪Von Hippel-Lindau(VHL)基因进行了基因敲低研究,以便为建立VHL疾病模型猪奠定基础.研究首先通过 3'RACE和5'RACE克隆得到猪VHL基因cDNA全长序列(2 725 bp),Real-time PCR结果表明VHL基因广泛表达于猪的各种组织器官,其中在肾上腺、肝脏、胰腺、心脏和睾丸等组织器官高量表达.进一步在猪iPS细胞中对5条干扰片段进行筛选,获得2条高效的干扰片段,干扰效率分别达到72％(P=0.0012)和64％ (P＜ 0.01).以稳定干扰VHL基因的猪胎儿成纤维细胞为核供体,构建克隆胚胎,结果表明,克隆胚胎的发育能力与对照组相比没有明显差异,而且在克隆囊胚中VHL基因的干扰效率达到71％ (P＜ 0.01).综上所述,文中获得了猪VHL基因全长序列并获得该基因稳定敲低的猪细胞和胚胎,从而为VHL疾病模型猪的构建奠定了良好的基础.     

Conversion of glycerol to 1,3-dihydroxyacetone by glycerol dehydrogenase co-expressed with an NADH oxidase for cofactor regeneration

Objectives

To investigate the efficiency of a cofactor regeneration enzyme co-expressed with a glycerol dehydrogenase for the production of 1,3-dihydroxyacetone (DHA).

Results

In vitro biotransformation of glycerol was achieved with the cell-free extracts containing recombinant GlyDH (glycerol dehydrogenase from Escherichia coli), LDH (lactate dehydrogenase form Bacillus subtilis) or LpNox1 (NADH oxidase from Lactobacillus pentosus), giving DHA at 1.3 g l^?1 (GlyDH/LDH) and 2.2 g l^?1 (GlyDH/LpNox1) with total turnover number (TTN) of NAD⁺ recycling of 6039 and 11100, respectively. Whole cells of E. coli (GlyDH–LpNox1) co-expressing both GlyDH and LpNox1 were constructed and converted 10 g glycerol l^?1 to DHA at 0.2–0.5 g l^?1 in the presence of zero to 2 mM exogenous NAD⁺. The cell free extract of E. coli (GlyDH–LpNox) converted glycerol (2–50 g l^?1) to DHA from 0.5 to 4.0 g l^?1 (8–25 % conversion) without exogenous NAD⁺.

Conclusions

The disadvantage of the expensive consumption of NAD⁺ for the production of DHA has been overcome.

     

Ced-9 inhibits Al-induced programmed cell death and promotes Al tolerance in tobacco

Our previous data showed that apoptotic suppressors inhibit aluminum (Al)-induced programmed cell death (PCD) and promote Al tolerance in yeast cells, however, very little is known about the underlying mechanisms, especially in plants. Here, we show that the Caenorhabditis elegans apoptotic suppressor Ced-9, a Bcl-2 homologue, inhibited both the Al-induced PCD and Al-induced activity of caspase-like vacuolar processing enzyme (VPE), a crucial executioner of PCD, in tobacco. Furthermore, we show that Ced-9 significantly alleviated Al inhibition of root elongation, decreased Al accumulation in the root tip and greatly inhibited Al-induced gene expression in early response to Al, leading to enhancing the tolerance of tobacco plants to Al toxicity. Our data suggest that Ced-9 promotes Al tolerance in plants via inhibition of Al-induced PCD, indicating that conserved negative regulators of PCD are involved in integrated regulation of cell survival and Al-induced PCD by an unidentified mechanism.