Disruption of Spectrin-Like Cytoskeleton in Differentiating Keratinocytes by PKCδ Activation Is Associated with Phosphorylated Adducin

Spectrin is a central component of the cytoskeletal protein network in a variety of erythroid and non-erythroid cells. In keratinocytes, this protein has been shown to be pericytoplasmic and plasma membrane associated, but its characteristics and function have not been established in these cells. Here we demonstrate that spectrin increases dramatically in amount and is assembled into the cytoskeleton during differentiation in mouse and human keratinocytes. The spectrin-like cytoskeleton was predominantly organized in the granular and cornified layers of the epidermis and disrupted by actin filament inhibitors, but not by anti-mitotic drugs. When the cytoskeleton was disrupted PKCδ was activated by phosphorylation on Thr505. Specific inhibition of PKCδ(Thr505) activation with rottlerin prevented disruption of the spectrin-like cytoskeleton and the associated morphological changes that accompany differentiation. Rottlerin also inhibited specific phosphorylation of the PKCδ substrate adducin, a cytoskeletal protein. Furthermore, knock-down of endogenous adducin affected not only expression of adducin, but also spectrin and PKCδ, and severely disrupted organization of the spectrin-like cytoskeleton and cytoskeletal distribution of both adducin and PKCδ. These results demonstrate that organization of a spectrin-like cytoskeleton is associated with keratinocytes differentiation, and disruption of this cytoskeleton is mediated by either PKCδ(Thr505) phosphorylation associated with phosphorylated adducin or due to reduction of endogenous adducin, which normally connects and stabilizes the spectrin-actin complex

An endogenous nanomineral chaperones luminal antigen and peptidoglycan to intestinal immune cells.

In humans and other mammals it is known that calcium and phosphate ions are secreted from the distal small intestine into the lumen. However, why this secretion occurs is unclear. Here, we show that the process leads to the formation of amorphous magnesium-substituted calcium phosphate nanoparticles that trap soluble macromolecules, such as bacterial peptidoglycan and orally fed protein antigens, in the lumen and transport them to immune cells of the intestinal tissue. The macromolecule-containing nanoparticles utilize epithelial M cells to enter Peyer's patches, small areas of the intestine concentrated with particle-scavenging immune cells. In wild-type mice, intestinal immune cells containing these naturally formed nanoparticles expressed the immune tolerance-associated molecule 'programmed death-ligand 1', whereas in NOD1/2 double knockout mice, which cannot recognize peptidoglycan, programmed death-ligand 1 was undetected. Our results explain a role for constitutively formed calcium phosphate nanoparticles in the gut lumen and show how this helps to shape intestinal immune homeostasis

The solubility of palladium in chloride solutions and the distribution of platinum, palladium, and related elements in hydrothermal mineralization

Online access for this thesis was created in part with support from the Institute of Museum and Library Services (IMLS) administered by the Nevada State Library, Archives and Public Records through the Library Services and Technology Act (LSTA). To obtain a high quality image or document please contact the DeLaMare Library at https://unr.libanswers.com/ or call: 775-784-6945.As many as 87 occurrences of platinum group elements (PGE) enrichment in hydrothermal systems are known worldwide. When compared with the occurrence and distribution of other metals commonly found in hydrothermal systems, this number is very small, compatible with the perceived rarity of PGE. That Pd is the most abundant of the PGE in hydrothermal systems pervades the literature, although it is not strictly true. In order to define the character of PGE hydrothermal mineralization and enhance the understanding of the genesis of these occurrences this research focuses upon three critical areas: 1) the conditions under which Pd is soluble in aqueous fluids, and therefore available for concentration into hydrothermal lodes. Experiments to determine the solubility of Pd in NaCl solutions were undertaken, and this was augmented by a thorough review and synthesis of experimental and theoretical studies of the solubility of PGE in a variety of solutions reported in the literature, 2) a literature compilation and synthesis was completed for the better studied occurrences, and 3) substantial new geochemical data were generated for PGE and related elements in a number of different classes of mineralization. From this broad analysis of the character of this type of hydrothermal mineralization, an understanding of the genesis of these occurrences is developed. Palladium solubility as high as 70 ppm was measured in 3m NaCl solution at a pH of 1.5, temperature of 300°C, and f02 = Ni-NiO. Solubility is retrograde with respect to temperature over the range 300 to 700°C and is higher in solutions of acidic pH and high salinity. Salinities of greater than 3m have been reported m fluid inclusions from some of the better studied hydrothermal PGE occurrences. Theoretical studies indicate that Pt solubility and behavior is similar to that of Pd but that Pd may be somewhat more soluble than Pt. Platinum and Pd are also soluble m bisulfide solutions under conditions of alkaline pH, dissolved S of > 0.001m, and f02 low enough to prevent the oxidation S to sulfite and sulfate; temperature dependence may be either prograde or retrograde. They are also soluble as ammine complexes in solutions of high dissolved NH3, alkaline pH, and f02 low enough to prevent the oxidation of ammonia to nitrite and nitrate. Geologic relationships indicate that the presence of appropriate source rocks may be of fundamental importance in permitting the enrichment of PGE in hydrothermal mineralization. Both mafic/ultramafic rocks and black shales may be suitable source rocks from which to mobilize the PGE. In hydrothermal mineralization Pt/Pd varies from 1. Although a close relationship between PGE and Cu is widely observed, PGE may not be closely related to Cu rich zones in individual occurrences that exhibit strong metal zoning. There is, however, a persistent and close spatial relationship between PGE enrichment and enrichment in one or more of the metalloids As, Sb, Se, Te, and/or Bi. Only the layered mafic intrusions are known to host mineralization which is mined only for PGE and are very large resources. Few occurrences outside the layered mafic intrusions represent large resources of (byproduct) PGE

A Rapamycin-Based GMP-Compatible Process for the Isolation and Expansion of Regulatory T Cells for Clinical Trials

The concept of regulatory T cell (Treg)-based immunotherapy has enormous potential for facilitating tolerance in autoimmunity and transplantation. Clinical translation of Treg cell therapy requires production processes that satisfy the rigors of Good Manufacturing Practice (GMP) standards. In this regard, we report our findings on the implementation of a robust GMP compliant process for the ex vivo expansion of clinical grade Tregs, demonstrating the feasibility of this developed process for the manufacture of a final product for clinical application. This Treg isolation procedure ensured the selection of a pure Treg population that underwent a 300-fold expansion after 36 days of culture, while maintaining a purity of more than 75% CD4+CD25+FOXP3+ cells and a suppressive function of above 80%. Furthermore, we report the successful cryopreservation of the final product, demonstrating the maintenance of phenotype and function. The process outlined in this manuscript has been implemented in the ONE study, a multicenter phase I/IIa clinical trial in which cellular therapy is investigated in renal transplantation