Characterization and Quantification of Selenoprotein P: Challenges to Mass Spectrometry

Selenoprotein P (SELENOP) is an emerging marker of the nutritional status of selenium and of various diseases, however, its chemical characteristics still need to be investigated and methods for its accurate quantitation improved. SELENOP is unique among selenoproteins, as it contains multiple genetically encoded SeCys residues, whereas all the other characterized selenoproteins contain just one. SELENOP occurs in the form of multiple isoforms, truncated species and post-translationally modified variants which are relatively poorly characterized. The accurate quantification of SELENOP is contingent on the availability of specific primary standards and reference methods. Before recombinant SELENOP becomes available to be used as a primary standard, careful investigation of the characteristics of the SELENOP measured by electrospray MS and strict control of the recoveries at the various steps of the analytical procedures are strongly recommended. This review critically discusses the state-of-the-art of analytical approaches to the characterization and quantification of SELENOP. While immunoassays remain the standard for the determination of human and animal health status, because of their speed and simplicity, mass spectrometry techniques offer many attractive and complementary features that are highlighted and critically evaluated.     

Structural mobility of molecular bottle-brushes investigated by NMR relaxation dynamics

The structural mobility of monomeric units of molecular bottle-brushes was studied by a systematic evaluation of NMR relaxation dynamics. The spin-spin relaxation time (T₂) was determined by Carr-Purcell-Meiboom-Gill (CPMG) NMR spectroscopic measurements. T₂ for protons that reside on the exterior and interior of the bottle-brush macromolecules varied with the grafting density and side chain length in bottle-brush copolymers. Poly((2-(2-bromopropionyloxy)ethyl methacrylate-stat-methyl methacrylate)-graft-butyl acrylate) (poly((BPEM-stat-MMA)-graft-PBA) was studied as a model brush copolymer. The T₂ values for protons of MMA units in the brush backbone significantly decreased with increasing side chain length and grafting density of PBA. The mobility and relaxation times T₂ for the side chain PBA protons decreased with grafting density. However, after initial increase, the relaxation times eventually decreased with PBA side chain length.     

Determination of Sensory Thresholds of Selected Calcium Salts and Formulation of Calcium-fortified Pocket-type Flat Bread

Pita bread loaves were prepared from flours fortified with calcium carbonate, calcium sulfate, and tricalcium dicitrate at 8 ascending levels to provide ranges of 800 to 2500, 700 to 1500, and 400 to 2000 mg of added Ca/100 g flour, respectively. The detection thresholds of calcium salts in pita bread were determined by the 3‐alter‐native forced choice (3‐AFC) test and construction of dose‐response curves. Detection thresholds determined by calculating geometric mean of individual best estimate thresholds, using criterion of 50%‐above‐chance and probit analysis of 3‐AFC data, were in the middle region of calcium concentrations. Analysis of dose‐response curves yielded values for thresholds outside the range of surveyed calcium concentrations. The detection threshold of CaSO₄ (2724 mg/100 g) in pita bread was significantly higher (P < 0.01) than those of calcium carbonate (1984 mg/100 g) and tricalcium dicitrate (2132 mg/100 g). Calcium‐fortified pita bread was similar (P < 0.01) to its regular counterpart when formulated to contain 1254.6, 1772.5, or 1155 mg/100 g of CaCO₃, CaSO₄, or tricalcium dicitrate, respectively. At the indicated levels of fortification, calcium‐fortified pita bread is expected to provide between 61% and 126.5% of the recommended daily intake for calcium for Middle Eastern populations.     

Protein Arginine Methyltransferase (PRMT) Inhibitors—AMI-1 and SAH Are Effective in Attenuating Rhabdomyosarcoma Growth and Proliferation in Cell Cultures

Rhabdomyosarcoma (RMS) is a malignant soft tissue cancer that develops mostly in children and young adults. With regard to histopathology, four rhabdomyosarcoma types are distinguishable: embryonal, alveolar, pleomorphic and spindle/sclerosing. Currently, increased amounts of evidence indicate that not only gene mutations, but also epigenetic modifications may be involved in the development of RMS. Epigenomic changes regulate the chromatin architecture and affect the interaction between DNA strands, histones and chromatin binding proteins, thus, are able to control gene expression. The main aim of the study was to assess the role of protein arginine methyltransferases (PRMT) in the cellular biology of rhabdomyosarcoma. In the study we used two pan-inhibitors of PRMT, called AMI-1 and SAH, and evaluated their effects on proliferation and apoptosis of RMS cells. We observed that AMI-1 and SAH reduce the invasive phenotype of rhabdomyosarcoma cells by decreasing their proliferation rate, cell viability and ability to form cell colonies. In addition, microarray analysis revealed that these inhibitors attenuate the activity of the PI3K-Akt signaling pathway and affect expression of genes related to it.     

Are toxicological responses in laboratory (inbred) zebrafish representative of those in outbred (wild) populations? - A case study with an endocrine disrupting chemical

Laboratory animals tend to be more inbred and less genetically diverse than wild populations, and thus may differ in their susceptibility to chemical stressors. We tested this hypothesis by comparing the responses of related inbred (theoretical inbreeding F(IT) = n + 0.25) and outbred (F(IT) = n) zebrafish (Danio rerio) WIK/Wild family lines to an endocrine disrupting chemical, clotrimazole. Exposure of inbred and outbred zebrafish to 2.9 μg clotrimazole/L had no effect on survival, growth, or gonadal development. Exposure of both lines to 43.7 μg clotrimazole/L led to male-biased sex ratios compared with controls (87% versus 55% and 92% vs 64%, for inbred and outbred males, respectively), advanced germ cell development, and reduced plasma 11-ketotestosterone concentrations in males. However, outbred males (but not inbred males) developed testis that were more than twice the weight of controls, which corresponded with a proliferation of Leydig cells and maintenance of the expression (rather than down-regulation occurring in inbreds) of gonadal aromatase (cyp19a1a) and insulin-like growth factor (igf1). Our results illustrate that the effects of an endocrine disrupting chemical (clotrimazole) on some end points (here testis development) can differ between inbred and outbred zebrafish. This highlights the need for reporting pedigree/genetic information and consistency in the responses of laboratory animals (e.g., by using model compounds as positive controls).     

新婚10月，新郎“装备”

10月是结婚的高峰期很多新人都会在这个美丽的季节踏上红毯。人们普遍关注的是新娘的服饰与妆容，但是现在随着生活水平的提高．思想观念的改变．让许多男人也越来越注意自己的“面子”问题．特别是在自己结婚前夕。