An insulin receptor mutant (Asp707 → Ala), involved in leprechaunism, is processed and transported to the cell surface but unable to bind insulin

We have identified a homozygous mutation near the carboxyl terminus of the insulin receptor (IR) α subunit from a leprechaun patient, changing Asp707 into Ala. Fibroblasts from this patient had no high affinity insulin binding sites. To examine the effect of the mutation on IR properties, the mutant IR was stably expressed in Chinese hamster ovary cells. Western blot analysis and metabolic labeling showed a normal processing of the mutant receptor to α and β subunits. No increase in high affinity insulin binding sites was observed on Chinese hamster ovary cells expressing the mutant receptor, and also, affinity cross-linking of 125I- labeled insulin by disuccinimidyl suberate to these cells failed to label the mutant α subunit. Biotinylation of cell surface proteins by biotin succinimidyl ester resulted in efficient biotinylation of the mutant IR α and β subunits, showing its presence on the cell surface. On solubilization of the mutant insulin receptor in Triton X. 100-containing buffers, 125I- insulin was efficiently cross-linked to the receptor a subunit by disuccinimidyl suberate. These studies demonstrate that Ala707 IR is normally processed and transported to the cell surface and that the mutation distorts the insulin binding site. Detergent restores this site. This is an example of a naturally occurring mutation in the insulin receptor that affects insulin binding without affecting receptor transport and processing. This mutation points to a major contribution of the a subunit carboxyl terminus to insulin binding

Submucosal Gland Myoepithelial Cells Are Reserve Stem Cells That Can Regenerate Mouse Tracheal Epithelium

The mouse trachea is thought to contain two distinct stem cell compartments that contribute to airway repair-basal cells in the surface airway epithelium (SAE) and an unknown submucosal gland (SMG) cell type. Whether a lineage relationship exists between these two stem cell compartments remains unclear. Using lineage tracing of glandular myoepithelial cells (MECs), we demonstrate that MECs can give rise to seven cell types of the SAE and SMGs following severe airway injury. MECs progressively adopted a basal cell phenotype on the SAE and established lasting progenitors capable of further regeneration following reinjury. MECs activate Wnt-regulated transcription factors (Lef-1/TCF7) following injury and Lef-1 induction in cultured MECs promoted transition to a basal cell phenotype. Surprisingly, dose-dependent MEC conditional activation of Lef-1 in vivo promoted self-limited airway regeneration in the absence of injury. Thus, modulating the Lef-1 transcriptional program in MEC-derived progenitors may have regenerative medicine applications for lung diseases

Validation of an automated assay for the measurement of cupric reducing antioxidant capacity in serum of dogs

BACKGROUND: The objective of the present study was to optimize and validate an automated method to assess the total antioxidant capacity (TAC) in serum of dogs using the cupric reducing antioxidant capacity (CUPRAC) methodology (TAC(c)) with bathocuproinedisulfonic acid disodium salt as chelating agent, evaluating also possible variations due to the use of two different automated analyzers. The method is based on the reduction of Cu(2+) into Cu(1+) by the action of the non-enzymatic antioxidants that are present in the sample. RESULTS: Imprecision was low in both apparatus utilized, and the results were linear across serial Trolox and canine serum samples dilutions. Lipids did not interfere with the assay; however, hemolysis increased the TAC(c) concentrations. When TAC(c) concentrations were determined in ten healthy (control) dogs and in twelve dogs with inflammatory bowel disease (IBD), dogs with IBD had lower TAC(c) concentrations when compared with the healthy dogs. CONCLUSIONS: The method validated in this paper is precise, simple, and fast and can be easily adapted to automated analyzers

Detrital zircon age and provenance constraints on late Paleozoic ice-sheet growth and dynamics in Western and Central Australia

U–Pb dating and Hf-isotope provenance analysis of detrital zircons from the glaciogenic lower Permian Grant Group of the Canning Basin indicate sources principally from basement terranes in central Australia, with subordinate components from terranes to the south and north. Integrating these data with field outcrop and subsurface evidence for ice sheets, including glacial valleys and striated pavements along the southern and northern margins of the basin, suggests that continental ice sheets extended over several Precambrian upland areas of western and central Australia during the late Paleozoic ice age (LPIA). The youngest zircons constrain the maximum age for contemporaneous ice sheet development to the late Carboniferous (Kasimovian), whereas palynology provides a minimum age of early Permian (Asselian–Sakmarian). Considering the palynological age of the Grant Group within the context of regional and global climate proxies, the main phase of continental ice sheet growth was possibly in the Ghzelian–Asselian. The presence of ice sheets older than Kasimovian in western and central Australia remains difficult to prove given a regional gap in deposition possibly covering the mid-Bashkirian to early Ghzelian within the main depocentres and even larger along basin margins, and the poor evidence for older Carboniferous glacial facies. There is also no evidence for extensive glacial facies younger than mid-Sakmarian in this region as opposed to eastern Australia where the youngest regional glacial phase was Guadalupian

A chelating ion exchanger for gallium recovery from alkaline solution using 5-palmitoyl-8-hydroxyquinoline immobilized on a nonpolar adsorbent

The recently developed method of gallium recovery from alkaline solution by alkanoyl oxine/chloroform extraction has been improved by immobilizing palmitoyl oxine on hydrophobic macroporous styrene-divinylbenzene copolymer Amberlite XAD-2 and passing the Ga-containing alkaline solution of pH 13.5 through the synthesized resin column. The developed column showed reasonable efficiency after successive passages, and the selectivity of Ga over Al was very high, suggesting the utilizibility of the method in Ga recovery from the basic aluminate liquor of the Bayer process. The Ga capacity of the oxine-based resin was 3.94 mu mol/g. Two mg Ga retained on 10 g resin could be eluted with 25 mL of 2 N HCl at a throughput rate of 2 mL/min. The developed process has prospective use in Ga separation from Al in a strongly alkaline solution

Modifier and surfactant dependence of gallium extraction by alkanoyl oxinates from basic solutions

The recently developed method of Ga extraction from basic aluminate solution (pH 13) using an alkanoyl oxine, i.e., myristoyl (M.Ox), palmitoyl (P.Ox), and stearoyl oxine (S.Ox), solution in chloroform has been improved by incorporating 2-hexanone as a modifier and cetyl trimethyl ammonium bromide (CTAB) as a surfactant into the extraction system. 2-Hexanone synergistically enhanced the extraction rate and Ga yield as its volume percentage was increased from 5 to 20% in the organic phase. Other oxygen-donor modifiers such as methyl isobutyl ketone (MIBK) and 1-hexanol, were not as effective as 2-hexanone. By using 10% (by volume) of 2-hexanone in chloroform solution, the order of Ga extraction efficiency for alkanoyl oxinates was M.Ox > P.Ox > S.Ox, i.e., extraction increased with decreasing length of the alkyl group of oxine. The ketone-added system showed satisfactory aluminum tolerance at pH 13, and enabled the shifting of the extraction pH to slightly higher values, which is beneficial for the process economies of Ga recovery from Bayer aluminate liquor. Use of CTAB as a cationic surfactant reduced the equilibration time and increased the Ga yield; the most pronounced effect occurred around its critical micelle concentration, indicative of micellar catalysis via interfacial mechanisms