MDR-like ABC transporter AtPGP4 is involved in auxin-mediated lateral root and root hair development

Previous data have suggested an involvement of MDR/PGP-like ABC transporters in transport of the plant hormone auxin and, recently, AtPGP1 has been demonstrated to catalyze the primary active export of auxin. Here we show that related isoform AtPGP4 is expressed predominantly during early root development. AtPGP4 loss-of-function plants reveal enhanced lateral root initiation and root hair lengths both known to be under the control of auxin. Further, atpgp4 plants show altered sensitivities toward auxin and the auxin transport inhibitor, NPA. Finally, mutant roots reveal elevated free auxin levels and reduced auxin transport capacities. These results together with yeast growth assays suggest a direct involvement of AtPGP4 in auxin transport processes controlling lateral root and root hair development.     

Hypotonic stress induces E-cadherin expression in cultured human keratinocytes

Human epidermis marks the interface between internal and external environments with the major task being to maintain body hydration. Alternating exposure of skin to a dry or humid environment is likely to cause changes in the epidermal water gradient resulting in osmotic alterations of epidermal keratinocytes. The present in vitro approach studied the effect of hypotonicity on cell-cell contact. It was demonstrated that hypotonic stress applied to human epithelial cells (HaCaT, A-431) induced upregulation of E-cadherin at both, the protein and mRNA level. 5'-deletional mutants of the E-cadherin promoter identified an element ranging from -53 to +31 that conveyed strong transactivation under hypotonic stress. In order to define relevant upstream regulators members of the MAP kinase family, the epidermal growth factor receptor (EGFR) and protein kinase B/Akt (PKB/Akt) were investigated. Hypotonic conditions led to a fast activation of ERK1/2, SAPK/JNK, p38, EGFR and PKB/Akt with distinct activation patterns. Experiments using specific inhibitors showed that p38 contributes to the E-cadherin transactivation under hypotonic conditions. Further upstream, adhesion was found to be a prerequisite for E-cadherin transactivation in this model. In summary, the present study provides evidence that E-cadherin is an osmo-sensitive gene that responds to hypotonic stress. The function of this regulation may be found in morphological changes induced by cell swelling. It is likely that induction of E-cadherin contributes to the stabilization between adjacent cells in order to withstand the physical forces induced by hypotonicity.     

Precise blood lead analysis using a combined internal standard and standard addition approach with disposable screen-printed electrodes

We report here a highly accurate and efficient method for blood lead analysis (BLA) through the use of a disposable electrode. A new type of mercury-plated preanodized screen-printed carbon electrode, together with a thallium(III) internal standard, simplifies calibration and gives easily quantifiable signals for accurate BLA. A preanodization procedure improves the preconcentration ability of the working electrode through the effect of lead (Pb) complexing with an electrogenerated surface functional group, mainly >C=O. Under optimized conditions, the ratio of the anodic stripping peak currents of Pb and Tl (i.e., i(Pb)/i(Tl)) is linear against [Pb] in the window of 1-300 ppb with a correlation coefficient and detection limit (signal-to-noise ratio=3) of 0.999 and 0.23 ppb, respectively. This approach was used to analyze blood samples (n=55) from workers at a local battery factory. The results are consistent with those obtained from graphite furnace atomic absorption spectroscopy and confirm the applicability of the proposed method.     

Estimation of diagnostic test characteristics and prevalence of Giardia duodenalis in dairy calves in Belgium using a Bayesian approach

A Bayesian approach was used to determine both the test properties of three diagnostic test procedures and the prevalence of Giardia duodenalis in dairy calves in Belgium. A cross-sectional survey was conducted in the province of East Flanders, Belgium. Between September 2001 and December 2003, a total of 100 farms were visited and faecal samples were obtained rectally from 499 calves aged from newborn to 70 days. Because there is no gold standard for the diagnosis of a G. duodenalis infection in dairy calves, a subset of 235 samples obtained on the first 50 farms, was examined using three different assays: microscopical examination, an immunofluorescence assay (IFA) and an antigen detecting Elisa (ELISA). Based on the results of these three tests, Bayesian analysis indicated that the prevalence of G. duodenalis in dairy calves was 0.19 (95% Confidence Interval: 0.11-0.28) and that ELISA (Sensitivity (Se) 0.89 and Specificity (Sp): 0.90) and IFA (Se: 0.77 and Sp: 0.95) were both sensitive and specific diagnostic techniques, whereas microscopical examination was less sensitive (Se: 0.56 and Sp: 0.87). The proportion of positive farms was estimated as 0.42 (0.24-0.62). The prevalence and the cyst excretion in calves from different age categories were based on data obtained by IFA on all 499 samples. The prevalence was highest among four to five week old calves and remained high among older calves up to 10 weeks, but was lower among calves before the age of two weeks. The number of excreted cysts was estimated by IFA and ranged from 100 to 1,040,000 cysts per gram faeces, with a mean of 3516 cysts per gram faeces. The intensity of excretion peaked among four-week-old calves and remained high among calves up to the age of eight weeks. This is the first known study to use Bayesian analysis to estimate the prevalence of G. duodenalis in the faeces of dairy calves and to estimate test characteristics of diagnostic assays used for the detection of G. duodenalis.     

Use of a fluorescent internal protein standard to achieve quantitative two-dimensional gel electrophoresis

2-DE is a powerful separation method for complex protein mixtures. However, large intergel variations in spot intensity limit its use for quantitative proteomics studies. To address this issue, we developed a fluorescent internal protein standard for use in 2-DE analysis. Protein samples are spiked with an Alexa-labeled internal standard (ALIS) prior to separation with 2-DE. Due to the high extinction coefficient of the Alexa-fluor, incorporation of 0.1% of total protein is sufficient to allow visualization of the internal standard yet low enough to avoid interference in subsequent quantification and identification steps. Following 2-DE, total proteins are visualized with fluorescent postelectrophoretic stains spectrally separated from ALIS. Four protein stains, Deep Purple, Sulforhodamine G, ruthenium II-tris(bathophenanthroline disulfonate) (RuTBS), and SYPRO Ruby, including improved purification and staining protocols for RuTBS and ten-fold dilutions of SYPRO Ruby were evaluated. All staining protocols were compatible with the ALIS method and had similar LODs (1-4 ng) and dynamic ranges (10(3)). ALIS is a powerful normalization method for quantitative 2-DE which avoids potential problems associated with dual spot migration patterns observed in the DIGE method. Furthermore, ALIS provides significantly improved normality in the distribution of spot abundance-variance compared to normalization through division by the total spot volume.     

野生稻种质资源描述规范和数据标准应用

论述了野生稻种质资源描述规范、数据标准和数据质量控制规范编制原则、适用范围及应用策略,对于促进全国野生稻种质资源的整合、规范鉴定评价,以及信息与实物共享机制与体系的建立,推动野生稻优异种质的研究与利用,变资源优势为经济优势都具有重要的现实意义。     

Chemical composition and rumen degradability of three corn hybrids treated with insecticides against the European corn borer (Ostrinia nubilalis)

This experiment determined the chemical composition, rumen degradability (aNDF in stalks and starch in kernels) and in vitro gas production of kernels from three corn hybrids treated (TT) or not treated (control, CTR) with insecticides against the European corn borer (ECB, Ostrinia nubilalis). Two whole-plant silage hybrids belonging to the FAO rating 600 and 700 maturity class (S600 and S700, respectively) and one selected for grain production (G600, FAO rating 600, Dekalb-Monsanto Agricoltura S.p.A., Lodi, Italy) were sown in two main plots (TT and CTR) of an experimental field. Two subsequent treatments of pyrethroids (25 and 1.2 g/ha of cyfluthrin and deltamethrin, respectively) were applied to the TT plots. The insecticide treatment reduced the number of damaged plants (4.5 broken plants/plot versus 0.3 broken plants/plot, P<0.01) and increased the total grain yield by 11% (13.8 t/ha versus 12.4 t/ha), while hybrids did not differ. ECB larvae which bored into the stalk tunnels modified the chemical composition of stalks and kernels. In stalks, total sugars content (i.e. glucose, fructose, sucrose) was about twice that in TT versus CTR plants (123 g/kg versus 60 g/kg DM, P<0.01), while aNDF content was higher in CTR stalks (765 versus 702 g/kg DM, P<0.01). DM degradability after 48 h of incubation of stalks was higher in TT than in CTR, both in vitro (0.360 versus 0.298, P<0.01) and in situ (0.370 versus 0.298, P<0.05), while there were no differences in aNDF degradability. Kernels from TT plots contained less DM (615 g/kg versus 651 g/kg, P<0.01) and more CP (84 g/kg and 78 g/kg DM, P<0.05) than those from CTR plots, while in situ rumen starch disappearance and in vitro gas production were similar. Corn hybrid selected for yield of grain (G600) differed from S600 and S700 due to a higher (P<0.01) content of aNDF, ADF and lignin(sa) in the stalks, and a higher starch content (696 g/kg versus 674 and 671 g/kg DM, P<0.01) and CP (87 g/kg versus 77 and 76 g/kg DM, P<0.05) in grain. The G600 hybrid produced stalks with a lower (P<0.01) aNDF rumen degradability than the S600 and S700.     

高效液相色谱法检测UV-1164的含量

建立了用高效液相色谱法测定UV-1164含量的方法,用标准品建立工作曲线,采用外标法定量。UV-1164在50mg.L-1~1000 mg.L-1范围内呈良好线性关系,r=0.999,方法的RSD是2.0%,平均回收率是99.8%。     

Plasmodium berghei (ANKA): infection induces CYP2A5 and 2E1 while depressing other CYP isoforms in the mouse liver

It has been reported that malaria infection impairs hepatic drug clearance and causes a down-regulation of CYP-mediated monooxygenase activities in rodents and humans. In the present study, we investigated the effects of Plasmodium berghei infection on the activity of liver monooxygenases in female DBA/2 and C57BL/6 mice. In both mouse strains, P. berghei infection decreased activities mediated by CYP1A (EROD: DBA/2 65.3%, C57BL/6 44.7%) and 2B (BROD: DBA/2 64.3%, C57BL/6 49.8%) subfamily isoforms and increased activities mediated by 2A5 (COH: DBA/2 182.4%, C57BL/6 148.5%) and 2E1 (PNPH: DBA/2 177.8%, C57BL/6 128.5%) isoforms as compared to non-infected controls. Since malaria infection also produced an increase in ALT (273.1%) and AST (354.1%) activities in the blood serum, our findings are consistent with the view that CYP2A5 activity is induced by liver injury. An almost generalized depression of CYP-mediated activities has been found with numerous infections and inflammatory stimuli but an induction of CYP2A5 had been previously noted only in some viral hepatitis and trematode (liver fluke) infections.     

A high standard metabolic rate constrains juvenile growth

The allocation of energy to various components of an individual's energy budget is often viewed as a competitive process. As such, a tradeoff may exist between production (growth) and maintenance metabolism. One view of a potential tradeoff, termed “the principle of allocation”, suggests that individuals with lower maintenance metabolic expenditures may have higher growth rates. To determine whether such a tradeoff exists, I analyzed the relationship between growth rate and maintenance metabolism of 225 juvenile snapping turtles housed in the laboratory. I measured growth from hatching to 6 months of age, and then measured oxygen consumption and calculated standard metabolic rate. Mean growth rate was 0.19 g d⁻ and mean standard metabolic rate (SMR) was 1.41 kJ d⁻. Maintenance metabolism and growth were negatively correlated after both were adjusted for body mass. The results support the “principle of allocation” theory: individuals with higher standard metabolic rates tended to have low growth rates.