Nonelectrolyte permeability of liposomes of hydroxyfatty acid-containing phosphatidylcholines

Two phosphatidylcholines containing hydroxylated fatty acids, 1-palmitoyl-2-[5-hydroxy-6,8,11,14-eicosatetraenoyl]-sn-glycero-3- phosphocholine (1-palm-2-5HETE PC) and 1-palmitoyl-2-[15(S)-hydroxy-5,8,11,13- eicosatetraenoyl]-sn-glycero-3-phosphocholine (1-palm-2-15HETE PC), and one phosphatidylcholine containing nonhydroxylated fatty acids, 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphocholine (1-palm-2-arach PC) were synthesized. Permeation of small nonelectrolytes (glycerol, 1,2-propanediol, urea, methylurea, propionamide and dimethylformamide) was assessed in multilamellar liposomes containing these synthetic PCs plus egg yolk phosphatidycholine (EPC) in the presence and absence of cholesterol. In liposomes containing 23% cholesterol, 69.3% EPC and 7.7% of either 1-palm-2-5HETE PC or 1-palm-2-15HETE PC the permeability to small nonelectrolytes was 60 to 400% greater than in liposomes containing 23% cholesterol and 77% EPC. The HETE-containing PCs also increased permeability in liposomes without cholesterol but the effects were less striking. Addition of the synthetic PCs did not affect the energy of activation of permeation.     

Lipoxygenase metabolites of arachidonic acid do not induce mucus secretion from rabbit intestinal goblet cells in vitro

Lipoxygenase metabolites of arachidonic acid are effective mucus secretagogues in the respiratory tract but their efficacy in the intestinal tract was unknown. Mucosal explants and sheets of epithelial cells isolated from rabbit small and large intestine were exposed to leukotrienes B4, C4, and D4 and monohydroxyeicosatetraenoic acids 5-HETE, 12-HETE, and 15-HETE. Light and electron microscopic inspection of goblet cells in treated tissues failed to detect evidence of recent compound exocytosis of mucin granules or other morphological evidence of secretory activity. These results indicate that lipoxygenase metabolites are not directly responsible for the increased mucus secretion observed in ulcerative colitis.     

ESTIMATING PUP PRODUCTION OF HARP SEALS, PAGOPHILUS GROENLANDICUS, IN THE NORTHWEST ATLANTIC

Photographic and visual aerial surveys to determine current pup production of Northwest Atlantic harp seals were conducted off Newfoundland and in the Gulf of St. Lawrence during March 1999-Photographic surveys were conducted on all whelping concentrations between 14 and 24 March, whereas a visual survey was made of the southern Gulf concentrations on 14 March. Pup production was estimated to be 739,100 (SE = 96,300, CV = 13.0%) at the Front, 82,600 (SE = 22,500, CV = 27.2%) in the northern Gulf, and 176,200 (SE = 25,400, CV = 14.4%) in the southern Gulf (Magdalen Island) for a total of 997,900 (SE = 102,100, 10.2%). Changes in aerial survey estimates indicate that pup production has increased since 1994. A new method to correct for the temporal change in the proportion of pups present on the ice was examined by fitting the percentage of pups observed in three age-dependent stages to a Normal distribution. The results were compared to those obtained from a more complex model used previously. The Simple model produced slightly higher, and hence more conservative, estimates of the proportion of births that had occurred before the time of the survey than the Complex model. When using the Simple model fewer assumptions regarding the start date of pupping and the proportion of older pups remaining on the ice were required, the herd had to be followed for a shorter period, and a more convenient means of calculating confidence limits was available.     

Ouabain-insensitive salt and water movements in duck red cells. II. Norepinephrine stimulation of sodium plus potassium cotransport

Catecholamines induce net salt and water movements in duck red cells incubated in isotonic solutions. The rate of this response is approximately three times greater than a comparable effect observed in 400 mosmol hypertonic solutions in the absence of hormone (W.F. Schmidt and T. J. McManus. 1977 a.J. Gen. Physiol. 70:59-79. Otherwise, these two systems share a great many similarities. In both cases, net water and salt movements have a marked dependence on external cation concentrations, are sensitive to furosemide and insensitive to ouabain, and allow the substitution of rubidium for external potassium. In the presence of ouabain, but the absence of external potassium (or rubidium), a furosemide-sensitive net extrusion of sodium against a large electrochemical gradient can be demonstrated. When norepinephrine-treated cells are incubated with ouabain and sufficient external sodium, the furosemide-sensitive, unidirectional influxes of both sodium and rubidium are half- maximally saturated at similar rubidium concentrations; with saturating external rubidium, the same fluxes are half-maximal at comparable levels of external sodium. In the absence of sodium, a catecholamine-stimulated, furosemide-sensitive influx of rubidium persists. In the absence of rubidium, a similar but smaller component of sodium influx can be seen. We interpret these results in terms of a cotransport model for sodium plus potassium which is activated by hypertonicity or norepinephrine. When either ion is absent from the incubation medium, the system promotes an exchange-diffusion type of movement of the co-ion into the cells. In the absence of external potassium, net movement of potassium out of the cell leads to a coupled extrusion of sodium against its electrochemical gradient.     

Esterification of monohydroxyfatty acids into the lipids of a macrophage cell line

Cells of a mouse macrophage-like tumor cell line, J774.2, were incubated with 0.6μM radiolabeled mono- and di-hydroxyfatty acids. Monohydroxyfatty acid products of the neutrophil and platelet lipoxygenase pathways (5-HETE, 15-HETE, and 12-HETE) were rapidly taken up (42–64% of the counts cell associated at 1 min) and esterified into triglycerides and phospholipids. 5-HETE and 12-HETE were found in triglycerides and distributed among phospholipid classes while 50% of added 15-HETE was esterified into phosphatidyl inositol. Treatment of phospholipids from cells incubated with 5-HETE, 12-HETE, and 15-HETE with phospholipase A₂ resulted in release of the respective monohydroxyfatty acid. HHT, a monohydroxyfatty acid product of the cyclooxygenase pathway, was taken up and esterified more slowly than the lipoxygenase products. In addition, HHT was not released when the phospholipids from cells incubated with HHT were treated with phospholipase A₂. LTB₄, a dihydroxyfatty acid product of neutrophil lipoxyegnase, was not taken up by J774.2 cells. The unique patterns of uptake and intracellular distribution of the different monohydroxyfatty acids suggests that the enzymes involved in the esterification of these compounds have substrate specificity and may also relate to the specific biologic effects of the compounds.     

Herbivoran impact on phytoplankton community structure

An enclosure experiment was conducted to assess the effects of a zooplankton elimination on the structure of a phytoplankton community. Phytoplankton biomass and production were higher in grazer-free enclosures, while the productivity per unit biovolume was lower. Exclusion of zooplankton favoured the majority of algal species, especially chrysophyceans (Dinobryon spp.) and the diatom Rhizosolenia, while mucilagineous green-algae were disfavoured. Middle sized algae (ESD 15–50 µm) and those with the largest Surface Area/Volume ratio were proportionally most favoured by the elimination of grazers.These differences in phytoplankton community structure are discussed in relation to effects of direct selective grazing and nutrient recycling by zooplankton. Some differences, as the immediate positive response of Dinobryon and Rhizosolenia, are probably caused by grazing release, while others, e.g. the response of mucilagineous species, might be caused by changed competitive relationships between the algae.     

MuPIT interactive: webserver for mapping variant positions to annotated, interactive 3D structures

Mutation position imaging toolbox (MuPIT) interactive is a browser-based application for single-nucleotide variants (SNVs), which automatically maps the genomic coordinates of SNVs onto the coordinates of available three-dimensional (3D) protein structures. The application is designed for interactive browser-based visualization of the putative functional relevance of SNVs by biologists who are not necessarily experts either in bioinformatics or protein structure. Users may submit batches of several thousand SNVs and review all protein structures that cover the SNVs, including available functional annotations such as binding sites, mutagenesis experiments, and common polymorphisms. Multiple SNVs may be mapped onto each structure, enabling 3D visualization of SNV clusters and their relationship to functionally annotated positions. We illustrate the utility of MuPIT interactive in rationalizing the impact of selected polymorphisms in the PharmGKB database, somatic mutations identified in the Cancer Genome Atlas study of invasive breast carcinomas, and rare variants identified in the exome sequencing project. MuPIT interactive is freely available for non-profit use at http://mupit.icm.jhu.edu.     

Characterization of a novel testicular form of human hormone-sensitive lipase

Hormone-sensitive lipase (HSL) is an esterase and lipase, which are essential for spermatogenesis. Two HSL mRNAs are expressed in human testis. A long form is encoded by a testis-specific exon and nine exons common to testis and adipocyte HSL. Here we show that the short-form 3.3-kb mRNA possesses a unique 5' end that is transcribed from a novel testis-specific exon. The corresponding protein is similar to the 775-amino-acid-long adipocyte HSL. Immunohistochemistry experiments on human testis sections revealed that the long form is strictly expressed in haploid germ cells whereas the short form is expressed in interstitial and tubular somatic cells as well as premeiotic germ cells.