Ivermectin and nodulisporic acid receptors in Drosophila melanogaster contain both gamma-aminobutyric acid-gated Rdl and glutamate-gated GluCl alpha chloride channel subunits

35S-labeled derivatives of the insecticides nodulisporic acid and ivermectin were synthesized and demonstrated to bind with high affinity to a population of receptors in Drosophila head membranes that were previously shown to be associated with a glutamate-gated chloride channel. Nodulisporic acid binding was modeled as binding to a single population of receptors. Ivermectin binding was composed of at least two kinetically distinct receptor populations, only one of which was associated with nodulisporic acid binding. The binding of these two ligands was modulated by glutamate, ivermectin, and antagonists of invertebrate gamma-aminobutyric acid (GABA)ergic receptors. Because solubilized nodulisporic acid and ivermectin receptors comigrated as 230-kDa complexes by gel filtration, antisera specific for both the Drosophila glutamate-gated chloride channel subunit GluCl alpha (DmGluCl alpha) and the GABA-gated chloride channel subunit Rdl (DmRdl) proteins were generated and used to examine the possible coassembly of these two subunits within a single receptor complex. DmGluCl alpha antibodies immunoprecipitated all of the ivermectin and nodulisporic acid receptors solubilized by detergent from Drosophila head membranes. DmRdl antibodies also immunoprecipitated all solubilized nodulisporic receptors, but only approximately 70% of the ivermectin receptors. These data suggest that both DmGluCl alpha and DmRdl are components of nodulisporic acid and ivermectin receptors, and that there also exists a distinct class of ivermectin receptors that contains the DmGluCl alpha subunit but not the DmRdl subunit. This co-association of DmGluCl alpha and DmRdl represents the first biochemical and immunological evidence of coassembly of subunits from two different subclasses of ligand-gated ion channel subunits.     

Classification of the insulin-like growth factor binding proteins into three distinct categories according to their binding specificities

Competitive binding experiments with insulin-like growth factor (IGF)-1, IGF-2 and des-(1-3)-IGF-1 have confirmed the interpretation based on limited amino-terminal sequence analysis that at least three types of IGF binding protein occur. In addition to the acid stable subunit of the large serum binding protein which exhibits des-(1-3)-IGF-1 binding only slightly less than IGF-1, the small IGF binding proteins can be separated into two classes based on differences in des-(1-3)-IGF-1 and IGF-2 binding potencies.     

Chromosomes of Syrphidae

In this report the karyotypes of 54 species of the tribe Milesiini and of four species of the tribe Myoleptini are described in detail with illustrations and idiograms. These species belong in the genera Lejota, Myolepta, Blera, Calliprobola, Criorhina, Hadromyia, Milesia, Somula, Sphecomyia, Spilomyia, Syritta, Temnostoma, Tropidia and Xylota. Six species have 2n = 8 chromosomes, 35 have 2n = 10 (including Xylota nemorum which has about 20 extra microchromosomes in some specimens), 15 have 2n = 12, one has 2n = 14, and Somula decora has 2n = 10 large chromosomes plus about eight microchromosomes. The mean total complement length (TCL) for 347 complements analysed in these tribes was 53.7 but there is great variation between TCL's of complements analysed even from a single fly. Karyotypes of species of Myolepta in the Myoleptini resemble in certain respects those of species of Tropidia in the Milesiini. Our observations support Currran's transfer of Lejota cyanea to the Milesiini. The 2n = 12 karyotypes of species of Blera, Criorhina, Lejota, Milesia, and to a lesser extent Sphecomyia, have some features in common. Spilomyia species have rather distinct 2n = 10 karyotypes. Certain species in Calliprobola, Syritta and Hadromyia are karyologically similar to some species of the genus Xylota in which species studied fall into fairly distinct karyological groups. These observations provide clear evidence of the accumulation of karyotypic variations in the origin of species in these two tribes.     

A scapular coordinate frame for clinical and kinematic analyses

The aim of this study was to define a body-fixed coordinate frame for the scapula that minimises axes variability and is closely related to the clinical frame of reference. Medical images of 21 scapulae were used to quantify 14 different axes from identifiable landmarks. The plane of the blade of the scapula was defined. The orientations of the quantified axes were calculated. The angular relationships between axes were quantified and applied to grade the sensitivity of each axis to inter-scapular variations in the others. The volume of data required to define an axis was noted for its dependency on pathology and the three criteria were weighted according to relative importance. The two axes with the highest weighting were applied to define a body-fixed Cartesian coordinate frame for the scapula. A least square medio-lateral line through the centre of the spine root was the most optimal axis. The plane formed by the spine root line and a least square line through the centre of the lateral border ridge was the most optimal scapular plane. This body-fixed Cartesian coordinate frame is closely aligned to the cardinal planes in the anatomical position and thus is a clinically applicable, specimen invariant coordinate frame that can be used in patient-specific kinematics modelling.     

Loss of TBL1XR1 Disrupts Glucocorticoid Receptor Recruitment to Chromatin and Results in Glucocorticoid Resistance in a B-Lymphoblastic Leukemia Model

Although great advances have been made in the treatment of pediatric acute lymphoblastic leukemia, up to one of five patients will relapse, and their prognosis thereafter is dismal. We have previously identified recurrent deletions in TBL1XR1, which encodes for an F-box like protein responsible for regulating the nuclear hormone repressor complex stability. Here we model TBL1XR1 deletions in B-precursor ALL cell lines and show that TBL1XR1 knockdown results in reduced glucocorticoid receptor recruitment to glucocorticoid responsive genes and ultimately decreased glucocorticoid signaling caused by increased levels of nuclear hormone repressor 1 and HDAC3. Reduction in glucocorticoid signaling in TBL1XR1-depleted lines resulted in resistance to glucocorticoid agonists, but not to other chemotherapeutic agents. Importantly, we show that treatment with the HDAC inhibitor SAHA restores sensitivity to prednisolone in TBL1XR1-depleted cells. Altogether, our data indicate that loss of TBL1XR1 is a novel driver of glucocorticoid resistance in ALL and that epigenetic therapy may have future application in restoring drug sensitivity at relapse.     

Indene bioconversion by a toluene inducible dioxygenase of Rhodococcus sp. I24

Rhodococcus sp. I24 can oxygenate indene via at least three independent enzyme activities: (i) a naphthalene inducible monooxygenase (ii) a naphthalene inducible dioxygenase, and (iii) a toluene inducible dioxygenase (TID). Pulsed field gel analysis revealed that the I24 strain harbors two megaplasmids of 340 and 50 kb. Rhodococcus sp. KY1, a derivative of the I24 strain, lacks the 340 kb element as well as the TID activity. Southern blotting and sequence analysis of an indigogenic, I24-derived cosmid suggested that an operon encoding a TID resides on the 340 kb element. Expression of the tid operon was induced by toluene but not by naphthalene. In contrast, naphthalene did induce expression of the nid operon, encoding the naphthalene dioxygenase in I24. Cell free protein extracts of Escherichia coli cells expressing tidABCD were used in HPLC-based enzyme assays to characterize the indene bioconversion of TID in vitro. In addition to 1-indenol, indene was transformed to cis-indandiol with an enantiomeric excess of 45.2% of cis-(1S,2R)-indandiol over cis-(1R,2S)-indandiol, as revealed by chiral HPLC analysis. The K_m of TID for indene was 380 M. The enzyme also dioxygenated naphthalene to cis-dihydronaphthalenediol with an activity of 78% compared to the formation of cis-indandiol from indene. The K_m of TID for naphthalene was 28 M. TID converted only trace amounts of toluene to 1,2-dihydro-3-methylcatechol after prolonged incubation time. The results indicate the role of the tid operon in the bioconversion of indene to 1-indenol and cis-(1S,2R)-indandiol by Rhodococcus sp. I24.