The effect of titanium dioxide nanoparticles on pulmonary surfactant function and ultrastructure

Background

Pulmonary surfactant reduces surface tension and is present at the air-liquid interface in the alveoli where inhaled nanoparticles preferentially deposit. We investigated the effect of titanium dioxide (TiO₂) nanosized particles (NSP) and microsized particles (MSP) on biophysical surfactant function after direct particle contact and after surface area cycling in vitro. In addition, TiO₂ effects on surfactant ultrastructure were visualized.

Methods

A natural porcine surfactant preparation was incubated with increasing concentrations (50-500 μg/ml) of TiO₂ NSP or MSP, respectively. Biophysical surfactant function was measured in a pulsating bubble surfactometer before and after surface area cycling. Furthermore, surfactant ultrastructure was evaluated with a transmission electron microscope.

Results

TiO₂ NSP, but not MSP, induced a surfactant dysfunction. For TiO₂ NSP, adsorption surface tension (γ_ads) increased in a dose-dependent manner from 28.2 ± 2.3 mN/m to 33.2 ± 2.3 mN/m (p < 0.01), and surface tension at minimum bubble size (γ_min) slightly increased from 4.8 ± 0.5 mN/m up to 8.4 ± 1.3 mN/m (p < 0.01) at high TiO₂ NSP concentrations. Presence of NSP during surface area cycling caused large and significant increases in both γ_ads (63.6 ± 0.4 mN/m) and γ_min (21.1 ± 0.4 mN/m). Interestingly, TiO₂ NSP induced aberrations in the surfactant ultrastructure. Lamellar body like structures were deformed and decreased in size. In addition, unilamellar vesicles were formed. Particle aggregates were found between single lamellae.

Conclusion

TiO₂ nanosized particles can alter the structure and function of pulmonary surfactant. Particle size and surface area respectively play a critical role for the biophysical surfactant response in the lung.     

Allergen particle binding by human primary bronchial epithelial cells is modulated by surfactant protein D

Background

Allergen-containing subpollen particles (SPP) are released from whole plant pollen upon contact with water or even high humidity. Because of their size SPP can preferentially reach the lower airways where they come into contact with surfactant protein (SP)-D. Our previous work demonstrated that SP-D increases the uptake of SPP by alveolar macrophages. In the present study, we investigated the uptake of SPP in human primary epithelial cells and the potential modulation by SP-D. The patho-physiological consequence was evaluated by measurement of pro-inflammatory mediators.

Methods

SPP were isolated from timothy grass and subsequently fluorescently labelled. Human primary bronchial epithelial cells were incubated with SPP or polystyrene particles (PP) in the presence and absence of surfactant protein D. In addition, different sizes and surface charges of the PP were studied. Particle uptake was evaluated by flow cytometry and confocal microscopy. Soluble mediators were measured by enzyme linked immunosorbent assay or bead array.

Results

SPP were taken up by primary epithelial cells in a dose dependent manner. This uptake was coincided with secretion of Interleukin (IL)-8. SP-D increased the fraction of bronchial epithelial cells that bound SPP but not the fraction of cells that internalized SPP. SPP-induced secretion of IL-8 was further increased by SP-D. PP were bound and internalized by epithelial cells but this was not modulated by SP-D.

Conclusions

Epithelial cells bind and internalize SPP and PP which leads to increased IL-8 secretion. SP-D promotes attachment of SPP to epithelial cells and may thus be involved in the inflammatory response to inhaled allergen.     

Biomarker discovery in heterogeneous tissue samples -taking the in-silico deconfounding approach

Background  

For heterogeneous tissues, such as blood, measurements of gene expression are confounded by relative proportions of cell types involved. Conclusions have to rely on estimation of gene expression signals for homogeneous cell populations, e.g. by applying micro-dissection, fluorescence activated cell sorting, or in-silico deconfounding. We studied feasibility and validity of a non-negative matrix decomposition algorithm using experimental gene expression data for blood and sorted cells from the same donor samples. Our objective was to optimize the algorithm regarding detection of differentially expressed genes and to enable its use for classification in the difficult scenario of reversely regulated genes. This would be of importance for the identification of candidate biomarkers in heterogeneous tissues.     

Estimation of breed contributions to present and future genetic diversity of 44 North Eurasian cattle breeds using core set diversity measures

Extinction of breeds threatens genetic diversity of livestock species. The need to conserve genetic diversity is widely accepted but involves in general two questions: (i) is the expected loss of diversity in a set of breeds within a defined future time horizon large enough to establish a conservation plan, and if so (ii) which breeds should be prioritised for such a conservation plan? The present study uses a marker assisted methodology to address these questions. The methodology combines core set diversity measures with a stochastic method for the estimation of expected future diversity and breed marginal diversities. The latter is defined as the change in the total diversity of all breeds caused by a one unit decrease in extinction probability of a particular breed. The stochastic method was validated by means of simulations. A large field data set consisting of 44 North Eurasian cattle breeds was analysed using simplified determined extinction probabilities. The results show that the expected loss of diversity in this set within the next 20 to 50 years is between 1 and 3% of the actual diversity, provided that the extinction probabilities which were used are approximately valid. If this loss is to be reduced, it is sufficient to include those three to five breeds with the highest marginal diversity in a conservation scheme.     

中国真蝽属二新种(半翅目,蝽科)

记述了蝽科真蝽属2新种:苍山真蝽Pentatoma cangshanensis sp.nov.和绿角真蝽Pentatoma viridicornuta sp.nov.,新种分别与近似种做了比较,并附鉴别特种图.模式标本保存于陕西师范大学动物研究所昆虫标本室.     

中国叶腿青蜂属一新种(膜翅目,青蜂科)

记述采自广西九万大山的青蜂科1新种:广西叶腿青蜂Loboscelidia guangxiensis sp.nov..模式标本保存于浙江大学寄生蜂标本室.广西叶腿青蜂,新种Loboscelidia guangxiensis sp.nov.(图1～6)新种与采自越南的亚细亚叶腿青蜂L.asiana Kimsey,1988近似,但其体色暗褐色、额突前观呈梯形、颈突侧观近直角弯曲、前翅缺M脉与cu-a脉.正模♂,广西九万大山,2003-07-31,王义平采,No.20037702.副模1♂,广西九万大山,2003-07-31,王义平采,No.20037720.     

三只羊岩溶峰丛洼地果-药复合栽培模式研究

岩溶峰丛洼地的耕地可分为3种类型：洼底平地、带状阶梯地和高坡石穴地。传统的玉米＋黄豆种植模式在三种类型耕地上的产值均很低,需要调整品种结构,改革种植模式。该文报道了基于三种耕地特点的四种果一药复合种植试验,即沙田柚＋绞股蓝、大果枇杷＋射干＋扶芳藤、无核黄皮＋射干＋扶芳藤、四月桃＋金银花。初步结果显示,四种模式均可大幅度提高耕地的产值,可作为岩溶峰丛洼地地区生态农业可持续发展的重要模式。     

抗欧亚活血丹凝集素特异性多克隆抗体的制备

欧亚活血丹外源凝集素(Gleheda)是分离自欧亚活血丹 (Glechoma hederacea) 叶片中的一种糖基化植物新蛋白. 如同其他糖基化蛋白,通过免疫学方法探测 Gleheda 的过程中通常受到一些不相干糖蛋白的妨碍,为此制定了抗 Gleheda 特异性多克隆抗体的纯化方案. 免疫血清蛋白经硫酸铵选择性沉淀后,分别以 Gleheda 和刺槐外源凝集蛋白 (RPA) 结合在 Sepharose 4B作为亲和配体,采用亲和层析法连续纯化 2 次,然后进一步采用离子交换层析 Q Fast Flow 提纯. 经每一步骤提纯得到的抗体组分对 Gleheda 的特异性,均同时采用双向免疫扩散检验和 Western blot 分析. 结果表明,以 Gleheda 为配体,亲和纯化制备得到的抗体组分对叶片粗提物中的许多植物 (糖) 蛋白仍然表现交叉反应. 为除去由植物糖蛋白中的聚糖所引起这些非特异性交叉反应抗体,接着以 RPA 为配体再次进行亲和纯化,Western blot 分析显示,抗体的特异性得到提高但并非除去了所有非特异性交叉反应的抗体. 最后进一步采用离子交换层析制备得到仅抗 Gleheda 蛋白的特异性抗体组分,此抗体组分适用于免疫探测研究. 该抗体纯化制备程序简易而高效,而且不需要昂贵的设备.     

横断山区伞形科4种7个居群植物的核型研究

对横断山区伞形科棱子芹属2种植物(松潘棱子芹Pleurospermum franchetianumHemsl.和西藏棱子芹Pleurospermum hookeriC.B.Clarke var.thomsoniiC.B.Clarke)和茴芹属2种植物(异叶茴芹Pimpinella diversi-foliaDC.和锐叶茴芹Pimpinella argutaDiels)共7个居群进行体细胞染色体数目观察和核型比较分析,结果表明,棱子芹属和茴芹属植物属内种间染色体基数存在差异,其中松潘棱子芹为2n=2x=18=16sm 2st,西藏棱子芹为2n=2x=22=16m 6sm;茴芹属光果组中锐叶茴芹为2n=2x=22=22m,毛果组中异叶茴芹为2n=18=18st或2n=18=2sm 16st.松潘棱子芹、西藏棱子芹、锐叶茴芹的染色体数目和核型均为首次报道,从而为棱子芹属和茴芹属的分类和演化研究提供细胞学依据.