Morphological changes in diabetic kidney are associated with increased O-GlcNAcylation of cytoskeletal proteins including α-actinin 4

Abstract Purpose The objective of the present study is to identify proteins that change in the extent of the modification with O-linked N-acetylglucosamine (O-GlcNAcylation) in the kidney from diabetic model Goto-Kakizaki (GK) rats, and to discuss the relation between O-GlcNAcylation and the pathological condition in diabetes. Methods O-GlcNAcylated proteins were identified by two-dimensional gel electrophoresis, immunoblotting and peptide mass fingerprinting. The level of O-GlcNAcylation of these proteins was examined by immunoprecipitation, immunoblotting and in situ Proximity Ligation Assay (PLA). Results O-GlcNAcylated proteins that changed significantly in the degree of O-GlcNAcylation were identified as cytoskeletal proteins (α-actin, α-tubulin, α-actinin 4, myosin) and mitochondrial proteins (ATP synthase β, pyruvate carboxylase). The extent of O-GlcNAcylation of the above proteins increased in the diabetic kidney. Immunofluorescence and in situ PLA studies revealed that the levels of O-GlcNAcylation of actin, α-actinin 4 and myosin were significantly increased in the glomerulus and the proximal tubule of the diabetic kidney. Immunoelectron microscopy revealed that immunolabeling of α-actinin 4 is disturbed and increased in the foot process of podocytes of glomerulus and in the microvilli of proximal tubules. Conclusion These results suggest that changes in the O-GlcNAcylation of cytoskeletal proteins are closely associated with the morphological changes in the podocyte foot processes in the glomerulus and in microvilli of proximal tubules in the diabetic kidney. This is the first report to show that α-actinin 4 is O-GlcNAcylated. α-Actinin 4 will be a good marker protein to examine the relation between O-GlcNAcylation and diabetic nephropathy.</p

Efficient Genetic Method for Establishing Drosophila Cell Lines Unlocks the Potential to Create Lines of Specific Genotypes

Analysis of cells in culture has made substantial contributions to biological research. The versatility and scale of in vitro manipulation and new applications such as high-throughput gene silencing screens ensure the continued importance of cell-culture studies. In comparison to mammalian systems, Drosophila cell culture is underdeveloped, primarily because there is no general genetic method for deriving new cell lines. Here we found expression of the conserved oncogene RasV12 (a constitutively activated form of Ras) profoundly influences the development of primary cultures derived from embryos. The cultures become confluent in about three weeks and can be passaged with great success. The lines have undergone more than 90 population doublings and therefore constitute continuous cell lines. Most lines are composed of spindle-shaped cells of mesodermal type. We tested the use of the method for deriving Drosophila cell lines of a specific genotype by establishing cultures from embryos in which the warts (wts) tumor suppressor gene was targeted. We successfully created several cell lines and found that these differ from controls because they are primarily polyploid. This phenotype likely reflects the known role for the mammalian wts counterparts in the tetraploidy checkpoint. We conclude that expression of RasV12 is a powerful genetic mechanism to promote proliferation in Drosophila primary culture cells and serves as an efficient means to generate continuous cell lines of a given genotype

Kinetic and thermodynamics studies of as (III) adsorption onto iron nanoparticles entrapped ca-alginate beads

ABSTRACT : Iron nanoparticles (Fe2O3) have been successfully entrapped in biopolymer viz. calcium (Ca)-alginate beads. The scanning electron microscopy images indicate that, the alginate gel acts as a bridge that binds the nanoparticles together. The adsorption of As (III) ions from the aqueous solution using Iron nanoparticle entrapped Ca-alginate as an adsorbent has been investigated. Kinetic studies with 0.50, 1.0, and 2.0 and 3.0 mg As (III)/ L indicates that 83-98% Arsenic removal was achieved with entrapped Fe2O3 nanoparticle as compared with bare Ca-alginate beads over a one hour period. The influences of the initial pH, temperature, contact time and dosage of the adsorbent on adsorption performance have been experimentally verified by column method. The equilibrium data were fitted to the Langmuir and Freundlich isotherm equations. From the effect of temperature, thermodynamic parameters like ΔG°, ΔH°, and ΔS° were calculated. The mechanism of adsorption for metal ions onto nanoparticle entrapped Ca-alginate was investigated by using the experimental results. Results suggested that Arsenic adsorption on iron nanoparticle entrapped Ca-alginate was spontaneous and endothermic process. The obtained results showed that iron oxide impregnated calcium alginate can be used to remove Arsenic from aqueous solutions even at low concentration. This work suggests that Fe2O3 nanoparticles offer a promising method for toxic heavy metal removal especially Arsenic, which is reported to causing severe health problems in the state of West Bengal and countries such as Bangladesh ,Taiwan , Mexico etc

Optimization of a high-voltage MOSFET in ultra-thin 14nm FDSOI technology

session 6: Low voltageInternational audienc

Probing the mobility of lithium in LISICON: Li<SUP>+</SUP>/H<SUP>+</SUP> exchange studies in Li<SUB>2</SUB>ZnGeO<SUB>4</SUB> and Li<SUB>2+2x</SUB>Zn<SUB>1-x</SUB>GeO<SUB>4</SUB>

We investigated Li+/H+ exchange in the lithium ion conductors (LISICONS) [Li2+2xZn1-xGeO4; x = 0.5 (I) and x = 0.75 (II)] and their parent, &#947;-Li2ZnGeO4. Facile exchange of approximately 2x lithium ions per formula unit occurs with both the LISICONS in dilute acetic acid, while the parent material does not exhibit an obvious Li+/H+ exchange under the same conditions. The results can be understood in terms of lithium ion distribution in the crystal structures: the parent Li2ZnGeO4, where all the lithium ions form part of the tetrahedral framework structure, does not exhibit a ready Li+/H+ exchange; LISICONS, where lithium ions are distributed between framework (tetrahedral) and nonframework sites, undergo a facile Li+/H+ exchange of the nonframework site lithium ions. Accordingly, Li+/H+ exchange in dilute aqueous acetic acid provides a convenient probe to distinguish between the mobile and the immobile lithium ions in lithium ion conductors