Conserved Extracellular Cysteines Differentially Regulate the Potentiation Produced by Zn2+ in Rat P2X4 Receptors

One feature of the amino acid sequence of P2X receptors identified from mammalian species, Xenopus laevis and zebrafish is the conservation of ten cysteines in the extracellular loop. Little information is available about the role of these conserved ectodomain cysteines in the function of P2X receptors. Here, we investigated the possibility that ten conserved cysteine residues in the extracellular loop of the rat P2X4 receptor may regulate zinc potentiation of the receptor using a series of individual cysteine to alanine point mutations and functional characterization of recombinant receptors expressed in Xenopus oocytes. For the C116A, C132A, C159A, C165A, C217A and C227A mutants, 10 µM zinc did not significantly affect the current activated by an EC40 concentration of ATP. By contrast, 5 µM zinc shifted the ATP concentration-response curve to the right in a parallel manner for both the C261A and C270A mutants and the magnitudes of those shifts were similar to that of the wildtype receptor. Interestingly, for the C126A and C149A mutants, 5 µM zinc potentiated ATP-activated current, but increased the maximal response to ATP by 90% and 81% respectively, without significantly changing the EC50 value of ATP. Thus, these results suggest that cysteines and disulfide bonds between cysteines are differentially involved in the potentiation of the rat P2X4 receptor by zinc

A novel MIPgene mutation associated with autosomal dominant congenital cataracts in a Chinese family

BACKGROUND: The major intrinsic protein gene (MIP), also known as MIP26 or AQP0, is a member of the water-transporting aquaporin family, which plays a critical role in the maintenance of lifelong lens transparency. To date, several mutations in MIP (OMIM 154050) have been linked to hereditary cataracts in humans. However, more pathogenic mutations remain to be identified. In this study, we describe a four-generation Chinese family with a nonsense mutation in MIP associated with an autosomal dominant congenital cataract (ADCC), thus expanding the mutational spectrum of this gene. METHODS: A large four-generation Chinese family affected with typical Y-suture cataracts combined with punctuate cortical opacities and 100 ethnically matched controls were recruited. Genomic DNA was extracted from peripheral blood leukocytes to analyze congenital cataract-related candidate genes. Effects of the sequence change on the structure and function of proteins were predicted by bioinformatics analysis. RESULTS: Direct sequencing of MIP in all affected members revealed a heterozygous nucleotide exchange c.337C>T predicting an arginine to a stop codon exchange (p.R113X). The substitution co-segregated well in all the affected individuals in the family and was not found in unaffected members or in the 100 unrelated healthy controls. Bioinformatics analysis predicted that the mutation affects the secondary structure and function of the MIP protein. CONCLUSIONS: We identified a novel mutation of MIP (p.R113X) in a Chinese cataract family. This is the first nonsense mutation of MIP identified thus far. This novel mutation is also the first disease-causing mutation located in the loop C domain of MIP. The results add to the list of mutations of the MIP linked to cataracts

A 2D-3D non-contact anthropometric method for daily dressing state-takes young Asian women as example

Taking young Asian women as research object, this paper is to develop a 2D-3D non-contact body measurement and calculation method for daily dressing state used to generate the CWH (chest, waist, and hip) circumferences suitable for garment practical applications. The general research approach is ‘analyzing related curves of human body, finding their fitting functions and length calculating formulas through mathematical analysis and measuring experiment with large number of samples – extracting effective feature points data in daily dressing state by dynamic and static experiments – calculating such dimensions as width and thickness after curve fitting – generating CWH circumferences via formulas and making comparison’. The numerical results validate the 2D-3D method presented in this paper as a useful and effective approach. Being different from other 3D non-contact measuring methods, the authors provide a new way to get the CWH measurements in the condition of daily natural dressing, which not only optimizes the non-contact anthropometrics theory, but also breaks the static measuring mode. Furthermore, the bringing forth of the concept of ‘daily dressing state’ and the laboratory experiments of this paper would be worthy for practical use