Expansion of the ISWI chromatin remodeler family with new active complexes

ISWI chromatin remodelers mobilize nucleosomes to control DNA accessibility. Complexes isolated to date pair one of six regulatory subunits with one of two highly similar ATPases. However, we find that each endogenously expressed ATPase co‐purifies with every regulatory subunit, substantially increasing the diversity of ISWI complexes, and we additionally identify BAZ2B as a novel, seventh regulatory subunit. Through reconstitution of catalytically active human ISWI complexes, we demonstrate that the new interactions described here are stable and direct. Finally, we profile the nucleosome remodeling functions of the now expanded family of ISWI chromatin remodelers. By revealing the combinatorial nature of ISWI complexes, we provide a basis for better understanding ISWI function in normal settings and disease.     

Effects on German cockroach nymphs of contact exposure to IGRs, singly and in combination

Late instar German cockroach male and female nymphs were exposed continuously for two weeks to surfaces treated with fenoxycarb, diflubenzuron, and pyriproxyfen, singly and in combination. Concentrations were determined that eliminated or nearly eliminated reproduction in matings with untreated mates, either through mortality, effects on reproduction, or a combination of mortality and sterility (no hatch). The major effect of fenoxycarb, pyriproxyfen, and pyriproxyfen plus fenoxycarb was on reproduction. The major effect of diflubenzuron was mortality. No hatch occurred in matings of females that were exposed to low concentrations of pyriproxyfen and fenoxycarb (2 ng/cm² and 6 ng/cm², respectively); sterility was incomplete when females were exposed to 600 ng/cm² of diflubenzuron. Mortality and sterility acted together to eliminate productive matings (matings that produced nymphs) when relatively high concentrations of diflubenzuron were combined with one or both of the other insect growth regulators (IGRs). In the triple combination, very small amounts of fenoxycarb and pyriproxyfen (total 1.1 ng/cm²) combined with 200 ng/cm² of diflubenzuron eliminated productive matings of treated females, but similar results with treated males were found only at higher concentrations of each IGR.     

Seasonal aspects of daily ration and diet of largemouth bass,Micropterus salmoides,with an evaluation of gastric evacuation rates

Synopsis Sample of stomach contents collected on 10 dates from May to October, 1978, were used to describe the diet and estimate the daily ration of subadult largemouth bass (primarily age-III fish) in Lake Rebecca, Minnesota. The method of Elliott & Persson (1978) was used to estimate daily ration. Data from sources in the literature were used to quantify gastric evacuation, which was found to be adequately described by an exponential decay model. The exponent of gastric evacuation increased exponentially with temperature. Seasonal changes in the diet with respect to composition, distribution of food among stomachs, and food particle size were reflected in the seasonal pattern of growth. Weight gain and the formation of scale annuli did not occur until the diet shifted from large bluegills and insects to age-0 largemouth bass and bluegills. Estimates of daily ration ranged from almost zero in mid-May and early June to over 5% in late August. The use of median weights of stomach contents was found to yield more meaningful estimates of the daily ration of individual bass than those based on means. Estimates based on medians were consistent with the observed pattern of growth and with information on maintenance rations and satiation levels. A growth-limiting lack of suitably-sized forage fish apparently occurred in the early part of the growing season.Paper Number 11,657, Scientific Journal Series, Minnesota Agricultural Experiment Station, St. Paul, MN. 55108.     

Strain differences in the response of German cockroaches (Dictyoptera: Blattellidae) to emulsifiable concentrates.

The response of a susceptible and two pyrethroid-resistant field strains of the German cockroach, Blattella germanica, to four emulsifiable concentrates (ECs)--Tempo (cyfluthrin), Demon (cypermethrin), Dursban (chlorpyrifos) LO, and XRM 5184 (chlorpyrifos)--was investigated. Application rates were 0.4, 0.1, 0.5, and 0.5%, respectively. Susceptible cockroaches avoided dried formulations of the pyrethroids Tempo and Demon, but resistant cockroaches did not; avoidance was greater with Demon than with Tempo. Filter papers freshly impregnated (wet) with Demon and Tempo flushed susceptible cockroaches (Demon greater than Tempo). Resistant cockroaches were flushed only by Demon, but less so than susceptible strain cockroaches. No mortality occurred in experiments with the pyrethroids. Dried formulations of Dursban LO and XRM 5184 had little, if any, repellency to nymphs of either the susceptible or resistant strains. Initially (1-3 h), most cockroaches settled on treated papers. Subsequent movement off treated papers was a step in the process leading to knockdown. At 24 h, mortality of susceptible cockroaches was 100% in the experiment with Dursban LO and 95% in the experiment with XRM 5184. Highest mortality in a resistant strain (40%) was in the contact experiment with Dursban LO. In contact repellency and flushing experiments, mortality was higher with Dursban LO than with XRM 5184. Neither chlorpyrifos formulation flushed cockroaches effectively. Mortality in flushing experiments was less than in the contact repellency experiments.     

Discovery of 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid diamides that increase CFTR mediated chloride transport

A series of 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid diamides that increase chloride transport in cells expressing mutant cystic fibrosis transmembrane conductance regulator (CFTR) protein has been identified from our compound library. Analoging efforts and the resulting structure-activity relationships uncovered are detailed. Compound potency was improved over 30-fold from the original lead, yielding several analogs with EC(50) values below 10nM in our cellular chloride transport assay.     

Temporal and spatial patterns of spawning site use by two cavity-spawning darters in a Wisconsin stream

Synopsis There was partial spatial and complete temporal overlap of spawning by Etheostoma flabellare and E. nigrum in Marsh Creek, Wisconsin. E. flabellare tended to spawn at shallower depths than E. nigrum. Both species spawned over a greater range of depths in an area with few spawning sites (cavities under rocks or logs) than in an area where spawning sites were abundant. In the laboratory E. flabellare were able to prevent E. nigrum from occupying cavities they would have otherwise occupied. E. nigrum appear to be competitively inferior to E. flabellare in terms of spawning-site acquisition and retention, but possess several attributes that may allow them to persist in the presence of the latter.     

Novel fucolipids accumulating in human adenocarcinoma. II. Selective isolation of hybridoma antibodies that differentially recognize mono-, di-, and trifucosylated type 2 chain

A series of glycolipids having the X determinant (Gal beta 1----4 [Fuc alpha----3]GlcNAc) at the terminus and a fucosyl alpha 1----3 residue at the internal GlcNAc residue have been isolated and characterized from tumor tissues (Hakomori, S., Nudelman, E., Levery, S.B., and Kannagi, R. (1984) J. Biol. Chem. 259, 4672-4680. A series of monoclonal antibodies that differentially recognize glycolipids with mono-, di-, and trifucosylated type 2 chain have been isolated and characterized. The antibody FH4 shows a remarkable preferential reactivity towards di-/or trifucosylated type 2 chain, i.e. it does not react with monofucosylated structures, including lactofucopentaosyl (III) ceramide (III3FucnLc4), monofucosyl neolactonorhexaosylceramide (y2, V3FucnLc6), and monofucosyl neolactonoroctaosylceramide (Z1, VII3FucnLc8), but reacts well with di- and trifucosylated type 2 chain structures such as difucosyl neolactonorhexaosylceramide (III3V3Fuc2nLc6) and trifucosyl neolactonoroctaosylceramide (III3V3VII3Fuc3nLc8). Two other monoclonal antibodies, FH5 and ACFH18, preferentially react with trifucosylated type 2 chain structure (III3V3VII3Fuc3nLc8), although cross-reactivity with difucosylated type 2 chain (III3V3Fuc2nLc6) was observed. They showed a minimal cross-reaction with monofucosylated type 2 chain. In contrast, the antibody FH1 does not react with III3FucnLc4 but reacts with V3FucnLc6, III3V3Fuc2nLc6, and III3V3VII3Fuc3nLc8. Two monoclonal antibodies, FH2 and FH3, do not discriminate among various glycolipids having fucosylated type 2 chain, and their reactivities are essentially similar to previously established antibodies directed to the X determinant, such as anti-SSEA-1, WGHS 29, VEP8 and 9, My-1, etc. This series of antibodies will be useful to detect the specific type of glycolipid with fucosylated type 2 chain accumulating in human cancer and in undifferentiated cells.