Range and magnitude of the steric pressure between bilayers containing phospholipids with covalently attached poly(ethylene glycol).

The interactive properties of liposomes containing phospholipids with covalently attached poly(ethylene glycol) (PEG-lipids) are of interest because such liposomes are being developed as drug delivery vehicles and also are ideal model systems for measuring the properties of surface-grafted polymers. For bilayers containing PEG-lipids with PEG molecular weights of 350, 750, 2000, and 5000, pressure-distance relations have been measured by X-ray diffraction analysis of liposomes subjected to known applied osmotic pressures. The distance between apposing bilayers decreased monotonically with increasing applied pressure for each concentration of a given PEG-lipid. Although for bilayers containing PEG-350 and PEG-750 the contribution of electrostatic repulsion to interbilayer interactions was significant, for bilayers containing PEG-2000 and PEG-5000 the major repulsive pressure between bilayers was a steric pressure due to the attached PEG. The range and magnitude of this steric pressure increased both with increasing PEG-lipid concentration and PEG size, and the extension length of the PEG from the bilayer surface at maximum PEG-lipid concentration depended strongly on the size of the PEG, being less than 35 A for PEG-750, and about 65 A for PEG-2000 and 115 A for PEG-5000. The measured pressure-distance relations have been modeled in terms of current theories (deGennes, 1987; Milner et al., 1988b) for the steric pressure produced by surface-grafted polymers, as modified by us to take into account the effects of polymer polydispersity and the possibility that, at low grafting densities, polymers from apposing bilayers surfaces can interpenetrate or interdigitate. No one theoretical scheme is sufficient to account for all the experimental results. However, for a given pressure regime, PEG-lipid size, and PEG-lipid surface density, the appropriately modified theoretical treatment gives a reasonable fit to the pressure-distance data.     

Stereospecific assignment of the NH2 resonances from the primary amides of asparagine and glutamine side chains in isotopically labeled proteins

An HMQC-based pulse scheme is presented for the stereospecific assignment of asparagineand glutamine side-chain amide protons. The approach makes use of the recently developedquantitative-J correlation spectroscopy [Bax, A. et al. (1994) Methods Enzymol., 239,79–105] to distinguish the E and Z primary amide protons and, as such, eliminates theneed for assignments derived from more time-consuming and potentially ambiguous NOEmethods. An application of this method to a uniformly 15N,13C-labeled cellulose-bindingdomain is presented. When used in combination with a NOESY-HSQC experiment, thepredominant 2 dihedral angles of two asparagine side chains in this protein can also bedefined.     

Disomic Thinopyrum intermedium addition lines in wheat with barley yellow dwarf virus resistance and with rust resistances.

Zhong 5 is a partial amphiploid (2n = 56) between Triticum aestivum (2n = 42) and Thinopyrum intermedium (2n = 42) carrying all the chromosomes of wheat and seven pairs of chromosomes from Th. intermedium. Following further backcrossing to wheat, six independent stable 2n = 44 lines were obtained representing 4 disomic chromosome addition lines. One chromosome confers barley yellow dwarf virus (BYDV) resistance, whereas two other chromosomes carry leaf and stem rust resistance; one of the latter also confers stripe rust resistance. Using RFLP and isozyme markers we have shown that the extra chromosome in the Zhong 5-derived BYDV resistant disomic addition lines (Z1, Z2, or Z6) belongs to the homoeologous group 2. It therefore carries a different locus to the BYDV resistant group 7 addition, L1, described previously. The leaf, stem, and stripe rust resistant line (Z4) carries an added group 7 chromosome. The line Z3 has neither BYDV nor rust resistance, is not a group 2 or group 7 addition, and is probably a group 1 addition. The line Z5 is leaf and stem rust resistant, is not stripe rust resistant, and its homoeology remains unknown.     

Interdigitated hydrocarbon chain packing causes the biphasic transition behavior in lipid/alcohol suspensions

It has been shown recently by Rowe ((1983) Biochemistry 22, 3299-3305) that ethanol has a 'biphasic' effect on the transition temperature (Tm) of phosphatidylcholine bilayers, reducing Tm at low concentrations but increasing Tm at high concentrations. Our X-ray diffraction data show that this reversal of Tm is a consequence of the induction of an unusual gel phase, where the lipid hydrocarbon chains from apposing monolayers fully interpenetrate or interdigitate. The properties of this interdigitated phase also explain the lipid chain length dependence of the reversal in the Tm versus ethanol concentration curves and the narrow width of the transition at high ethanol concentrations, as well as spectroscopic and calorimetric data from lipid suspensions containing other drugs such as methanol, benzyl alcohol, phenyl ethanol, and chlorpromazine.     

Avoidance response of Midge Larvae (Chironomus Tentans) to sediments containing heavy metals

Avoidance reactions of chironomid larvae to contaminated sediment taken from a heavy metal impacted lake were studied. Heavy metal levels in the test sediment ranged from background of 0.6 parts per million (ppm) cadmium, 77 ppm zinc and 17 ppm chromium to a maximum of 1,029 ppm cadmium, 17,262 ppm zinc and 2,106 ppm chromium. A linear relationship was established between cadmium and zinc levels in the sediment and avoidance by chironomids.An approximate threshold avoidance of metals in the sediment was determined to be between 213–422 ppm cadmium, 4385–8330 ppm zinc and 799–1513 ppm chromium.Supported by NIH Training Grant Number 5T01-ES00071 from National Institute of Environmental Health Sciences and in part by an NSF (RANN) Grant Number GI-35106.Purdue University Agricultural Experiment Station, Jour. No. 6474.     

Coordinate regulation by iron of the synthesis of phenolate compounds and three outer membrane proteins in Escherichia coli.

The biosynthesis of the low-molecular-weight iron carrier enterochelin and of three outer membrane polypeptides appears to be coordinately regulated by the amount of cell-associated iron in Escherichia coli K-12. Measurements of iron acquisition made throughout the growth cycle in iron-deficient media indicate that a very rapid accumulation of iron occurs in the first 2 h of growth; there is comparatively little iron uptake during exponential growth, which results in a gradual decrease in the cellular iron content with each generation. When this level falls below 400 ng of iron per mg (dry weight) of cells, there is a simultaneous onset of synthesis of the three outer membrane polypeptides and of enterochelin. This coordinate regulation was also observed in cells able to transport iron actively using only citrate as an iron-carrier.     

Somatostatin and insulin release from isolated rat pancreatic islets in response to D-glucose, l-leucine, alpha-ketoisocaproic acid or D-glyceraldehyde: evidence for a regulatory role of adenosine-3' ,5'-cyclic monophosphate.

Somatostatin and insulin release from isolated rat pancreatic islets was stimulated by glucose, leucine or α-ketoisocaproic acid. D-glyceraldehyde stimulated insulin release but diminished the secretion of somatostatin. Glucagon and theophylline amplified the glucose-induced somatostatin release.A regulatory role of the D-cell's adenylate cyclase/phosphodiesterase system for the release of somatostatin is suggested. Furthermore, stimulation as well as inhibition of somatostatin release might be of significance for the secretory function of the B-cell.     

Acetazolamide in prevention of acute mountain sickness: a double-blind controlled cross-over study.

Twenty-four amateur climbers took part in a double-blind controlled cross-over trial of acetazolamide versus placebo for the prevention of acute mountain sickness. They climbed Kilimanjaro (5895 m) and Mt Kenya (5186 m) in three weeks with five rest days between ascents. The severity of acute mountain sickness was gauged by a score derived from symptoms recorded daily by each subject. On kilimanjaro those taking acetazolamide reached a higher altitude (11 v 4 reached the summit) and had a lower symptom score than those taking placebo (mean 4.8 v 14.3). Those who had taken acetazolamide on Kilimanjaro maintained their low symptom scores while taking placebo on Mt Kenya (mean score 1.9), whereas those who had taken placebo on Kilimanjaro experienced a pronounced improvement when they took acetazolamide on Mt Kenya (mean score 2.5). Acute mountain sickness prevented one subject for completing either ascent. Acetazolamide was acceptable to 23 of the 24 subjects. Acetazolamide is recommended as an acceptable and effective prophylactic for acute mountain sickness.