Novel voltammetric biosensor for determining acrylamide in food samples

Recent findings showing that acrylamide is formed in heat-treated foods rich in asparagine and reducing sugars such as glucose, have accelerated the needs for the development of new analytical methods to determine this potential human carcinogen. Acrylamide forms adduct with hemoglobin (Hb) as a result of the reaction with the alpha-NH2 group of N-terminal valine of Hb. This interaction is the basis of a new voltammetric biosensor to detect acrylamide. The biosensor was constructed using a carbon-paste electrode modified with hemoglobin (Hb), which contains four prosthetic groups of heme--Fe(III). Such an electrode displays a reversible reduction/oxidation process of Hb-Fe(III)/Hb-Fe(II). Interaction between Hb and acrylamide was observed through decreasing of the peak current of Hb-Fe(III) reduction. The electrodes presented a very low detection limit (1.2 x 10(-10)M). The validation made in the matrix obtained by water extraction of potato chips showed that the electrodes presented are suitable for the direct determination of acrylamide in food samples.     

Analysis of chameleon sequences and their implications in biological processes

Chameleon sequences have been implicated in amyloid related diseases. Here we report an analysis of two types of chameleon sequences, chameleon-HS (Helix vs. Strand) and chameleon-HE (Helix vs. Sheet), based on known structures in Protein Data Bank. Our survey shows that the longest chameleon-HS is eight residues while the longest chameleon-HE is seven residues. We have done a detailed analysis on the local and global environment that might contribute to the unique conformation of a chameleon sequence. We found that the existence of chameleon sequences does not present a problem for secondary structure prediction programs, including the first generation prediction programs, such as Chou-Fasman algorithm, and the third generation prediction programs that utilize evolution information. We have also investigated the possible implication of chameleon sequences in structural conservation and functional diversity of alternatively spliced protein isoforms.     

Ultrastructure of alimentary tract formation in embryos of two insect species: Melasoma saliceti and Chrysolina pardalina (Coleoptera, Chrysomelidae)

Embryogenesis of the alimentary tract in two chrysomelid species (Chrysolina pardalina and Melasoma saliceti) is described. The embryonic development of both species lasts 7days at room temperature. Stomodaeum and proctodaeum invaginate at the anterior and posterior ends of the germ band. Together with the ectodermal tissue the endoderm cells also enter into the embryo. The anterior and posterior parts of the alimentary tract wedge into the yolk in the form of conical structures. The endodermal cells remain at the yolk surface and start migration over the yolk mass as two lateral bands of cells. The endoderm is always accompanied by mesoderm. On the fifth day of development the endodermal cells together with the mesoderm layer spread over the ventral and dorsal sides of the yolk mass and form the single layered primordium of the midgut epithelium. On the sixth day of development a basal lamina appears between the endoderm and the mesoderm cells and differentiation of both tissues starts. The endodermal epithelium cells change shape from flat to cuboidal and eventually into columnar. Mesoderm cells differentiate into muscle and tracheae. On the 7thday of development stomodaeum and proctodaeum become lined with cuticle and the midgut becomes covered with microvilli. The yolk cells populating the yolk mass do not contribute to midgut formation in the species studied.     

Physiological and antioxidant responses of two accessions of Arabidopsis thaliana in different light and temperature conditions

During their lifetime, plants need to adapt to a changing environment, including light and temperature. To understand how these factors influence plant growth, we investigated the physiological and antioxidant responses of two Arabidopsis accessions, Shahdara (Sha) from the Shahdara valley (Tajikistan, Central Asia) in a mountainous area and Lovvik‐5 (Lov‐5) from northern Sweden to different light and temperature conditions. These accessions originate from different latitudes and have different life strategies, both of which are known to be influenced by light and temperature. We showed that both accessions grew better in high‐light and at a lower temperature (16°C) than in low light and at 23°C. Interestingly, Sha had a lower chlorophyll content but more efficient non‐photochemical quenching than Lov‐5. Sha, also showed a higher expression of vitamin E biosynthetic genes. We did not observe any difference in the antioxidant prenyllipid level under these conditions. Our results suggest that the mechanisms that keep the plastoquinone (PQ)‐pool in more oxidized state could play a role in the adaptation of these accessions to their local climatic conditions.     

A thiol‐disulfide oxidoreductase of the Gram‐positive pathogen Corynebacterium diphtheriae is essential for viability,pilus assembly,toxin production and virulence

The Gram‐positive pathogen Corynebacterium diphtheriae exports through the Sec apparatus many extracellular proteins that include the key virulence factors diphtheria toxin and the adhesive pili. How these proteins attain their native conformations after translocation as unfolded precursors remains elusive. The fact that the majority of these exported proteins contain multiple cysteine residues and that several membrane‐bound oxidoreductases are encoded in the corynebacterial genome suggests the existence of an oxidative protein‐folding pathway in this organism. Here we show that the shaft pilin SpaA harbors a disulfide bond in vivo and alanine substitution of these cysteines abrogates SpaA polymerization and leads to the secretion of degraded SpaA peptides. We then identified a thiol‐disulfide oxidoreductase (MdbA), whose structure exhibits a conserved thioredoxin‐like domain with a CPHC active site. Remarkably, deletion of mdbA results in a severe temperature‐sensitive cell division phenotype. This mutant also fails to assemble pilus structures and is greatly defective in toxin production. Consistent with these defects, the ΔmdbA mutant is attenuated in a guinea pig model of diphtheritic toxemia. Given its diverse cellular functions in cell division, pilus assembly and toxin production, we propose that MdbA is a component of the general oxidative folding machine in C. diphtheriae.     

云南常见牛肝菌属真菌中汞含量及食用安全评价

汞(Hg)是危害人类健康的主要重金属元素之一,多数食用菌对Hg有很强的富集能力,测定食用菌中Hg含量,并对其进行食用安全性评价具有重要意义。采用冷原子吸收直接测汞仪系统测定85份云南常见牛肝菌属真菌菌盖、菌柄中总Hg含量;以同一牛肝菌子实体菌盖与菌柄总Hg含量比(Q(C/S))分析牛肝菌属真菌对Hg的富集特征;根据联合国粮农组织和世界卫生组织(FAO/WHO)现行每周Hg允许摄入量(provisional tolerable weekly intake,PTWI)标准和中国GB 2762—2012规定的食用菌中Hg限量标准评价样品的食用安全性。结果表明,菌盖和菌柄中总Hg含量分别在0.13~22.00、0.20~8.40 mg·kg-1DW,不同产地、种类及不同采集年份的样品中总Hg含量存在明显差异;同一牛肝菌菌盖、菌柄总Hg含量比(Q(C/S))在0.28~4.08,92%的样品Q(C/S)1,表明多数样品中菌盖对Hg的富集能力强于菌柄。根据GB 2762—2012规定的食用菌及其制品中总Hg含量限量标准(≤0.1 mg·kg-1),发现所有测试样品的总Hg含量均超标;根据PTWI标准,假设成年人(60 kg)每周食用300 g新鲜牛肝菌,则多数样品Hg摄入量低于允许摄入量,无安全风险,而少数样品Hg摄入量高于允许摄入量,食用有一定的潜在风险。同时,牛肝菌总Hg含量与种类、产地、采集时间等因素密切相关,采食及安全评价需综合考虑这些因素。