Sugar orthocarbonates

A simple procedure is described for preparing sugar orthocarbonates. It is based on treating the corresponding thionocarbonate in pyridine with cupric acetate and an alcohol, such as methanol, ethanol, or isopropyl alcohol. Treatment of 1,2:5,6-di-O-isopropylidene-D-mannitol 3,4-thionocarbonate with diols, such as 1,2-ethanediol, 1,2-propanediol, or 1,2:5,6-di-O-isopropylidene-D-mannitol, also gave orthocarbonates. Methyl thionocarbonate, S-methyl xanthate, and dithiobis(thioformate) derivatives of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose all gave the trimethyl orthocarbonate upon treatment with methanol in the presence of pyridine and cupric acetate. The structure of the orthocarbonates was proved by elemental analysis, n.m.r., and mass spectra, and by treatment with mild acid to form carbonates. Treatment of 1,2:5,6-di-O-isopropylidene-3-thio-D-altritol 3,4-thionocarbonate with methanol or ethanol gave the corresponding orthothiocarbonate, but on treatment with 1,2-ethanediol or with sodium ethoxide the 3,4-episulfide resulted.     

Fish cell lines: Two new cell lines derived from explants of trunk musculature ofCynoscion arenarius

Summary Two cell lines were established from explants of trunk musculature of healthy, males sand seatrout,Cynoscion arenarius. One of the lines, designated CyA-1, has been carried through 150 subcultures during 6 yr. The other, designated CyA-2, has been carried through 100 subcultures during 2 yr. Both lines grow well in L15 medium adjusted to 0.150M NaCl and supplemented with 10% fetal bovine serum. Optimal growth occurs at temperatures between 24 and 30°C. The species of origin of both lines was confirmed by a cytotoxic antibody dye exclusion test. The karyotype of CyA-1 has not yet stabilized, showing a modal chromosome number of 120 at Passage 9, 89 at Passage 63, and 79 at Passage 100. The karyotype of CyA-2 is rather stable, with a modal chromosome number of 47 at Passage 1 and 49 at Passage 100. Chromosome morphology of CyA-2 is homogeneous (small, acrocentric), whereas the chromosomes of CyA-1 show considerable size variation (with small chromosomes possibly formed from fragmentation of original structures). Both lines were found to be free of bacterial or fungal contamination. Both lines supported replication of lymphocystis virus strains isolated fromBairdiella chrysura (the silver perch) and fromMicropogon undulatus (the Atlantic croaker) but were refractory to 11 other viruses (4 from fish, 1 from amphibians, and 6 from mammals). This study was supported in part by the National Oceanographic and Atmospheric Administration, Office of Sea Grant No. 04-3-158-58.     

A genetic linkage map for Arctic char (Salvelinus alpinus): evidence for higher recombination rates and segregation distortion in hybrid versus pure strain mapping parents.

We constructed a genetic linkage map for Arctic char (Salvelinus alpinus) using two backcrosses between genetically divergent strains. Forty-six linkage groups (expected = 39-41) and 19 homeologous affinities (expected = 25) were identified using 184 microsatellites, 129 amplified fragment length polymorphisms (AFLPs), 13 type I gene markers, and one phenotypic marker, SEX. Twenty-six markers remain unlinked. Female map distance (9.92 Morgans) was substantially higher than male map distance (3.90 Morgans) based on the most complete parental information (i.e., the F1 hybrids). Female recombination rates were often significantly higher than those of males across all pairwise comparisons within homologous chromosomal segments (average female to male ratios within families was 1.69:1). The female hybrid parent had significantly higher recombination rates than the pure strain female parent. Segregation distortion was detected in four linkage groups (4, 8, 13, 20) for both families. In family 3, only the largest fish were sampled for genotyping, suggesting that segregation distortion may represent regions possessing influences on growth. In family 2, almost all cases showing segregation distortion involved markers in the female hybrid parent.     

Heat-shock protein expression in a perennial grass commonly associated with active geothermal areas in western North America

Heat-shock protein (hsp) expression in response to short- (hours) versus long-term (weeks) heat was examined in Dichanthelium lanuginosum. Expression of cytoplasmic class I small hsps (shsps) and hsp 101 was elicited by 2 h above 40°C, along with an increase over basal levels of hsp 70. Elevated levels of heat-induced shsp and hsp 101 persisted for 5–7 days after plants were returned to control temperatures. Protein extracts from roots exposed to 42°C for several weeks displayed increased levels of shsp but decreased hsp 101 levels. This decrease in hsp 101 did not occur in D. lanuginosum from normothermic environments.     

Possible role of transient electric fields in freezing-induced membrane destabilization

Summary Evidence is presented to support the hypothesis that electrical potentials generated during the freezing of aqueous solutions (the Workman-Reynolds effect) may contribute to the destabilization of the plasma membrane and cryoinjury of isolated protoplasts. Specifically. (1) electric potential diffrences of sufficient magnitude to cause lysis of the plasma membrane occur during the rapid freezing of isolated protoplasts suspended in sorbitol: (2) survival of protoplasts is inversely correlated with the magnitude of the potential difference and (3) cold acclimation increases the stability of the plasma membrane to applied electric fields. A discussion is given of the different physical phenomena though to be involved in the Workman-Reynolds effect. The basis equations for these phenomena are outlined.     

Characterization of molybdate-stabilized estrogen receptors by hydrophobic interaction HPLC: resolution of two 8S subunits

This report describes the separation and characterization of estrogen receptors (ER) according to their degree of hydrophobicity and surface charges. Molybdate-stabilized [3H]ER from rabbit uterine cytosol was sequentially purified by passage through a size-exclusion pre-column, an anion-exchange column, and a hydrophobic interaction column. With fresh cytosol, a major radioactive peak was eluted from the DEAE columns; a major peak and a minor, less hydrophobic, peak were eluted from the hydrophobic column. In contrast, ER from frozen cytosol showed one peak in the DEAE-column and exhibited four radioactive peaks in the hydrophobic column. By sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis, [3H] tamoxifen aziridine(TA)-labelled ER showed radioactive bands at 62 and 48 kd. The subunits which were characterized by these radioactive bands were successfully separated by the hydrophobic column; the more hydrophobic subunit corresponded to the 62 kd band. The HPLC-purified [3H]TA-labelled ER subunits sediment at a 7.4-8.5S region in a low-salt sucrose gradient. These results show that differential negative surface charge and hydrophobic areas exist in the holo-receptor and its subunits, and the hydrophobic interaction HPLC column separates the two major 8S steroid binding subunits of ER.