Changes in lipid composition of tritosomes during lysis

Membrane phospholipids and free fatty acids were analyzed after lysis of lysosomes which had engulfed Triton WR 1339 (tritosomes). Significant elevations of lysophosphatidyl choline and lysophosphatidyl ethanolamine were seen. Phosphatidyl choline, phosphatidyl ethanolamine and sphingomyelin decreased significantly. Triglyceride also fell. The released free fatty acids paralleled closely the liberation of β-glucuronidase from the tritosomes. Thus, lipases hydrolyzed endogenous phospholipids and neutral lipids of tritosomes during incubation at 37°C at pH 5.0 to produce significant elevations of lysophospholipids and free fatty acids. The known surface-active properties of these latter compounds raise the possibility of their participation in the process of lysis of the tritosomes.     

Migration inhibitory factor (MIF) and proliferative responses by human lymphocyte subpopulations separated by sheep erythrocyte rosette formation.

Human peripheral blood lymphocytes were separated by a combination of rosette formation with sheep erythrocytes and differential density centrifugation into subpopulations of rosette positive (T-enriched) cells and rosette negative (T depleted) cells. These were then tested in vitro for the production of macrophage migration inhibitory factor (MIF) and for incorporation of ³H-thymidine in response to specific antigens. Both T enriched and T depleted cell populations produced MIF but only T enriched cells exhibited significant antigen-induced ³H-thymidine incorporation. These findings using a T cell surface marker as the basis for cell separation, a technique which should not alter the B cell surface, confirm an earlier report in which human cells were separated on the basis of surface immunoglobulin, a B cell marker.     

A neural model of interactions subserving prey-predator discrimination and size preference in anuran amphibia

The model described is an extension of a previous model of the optic tectum (Arbib & Lara, 1982; Lara, Arbib & Cromarty, 1982; Lara & Arbib, 1982) and takes into consideration anatomical, physiological and behavioral studies in anurans, as well as earlier modelling efforts (Ewert & Von Seelen, 1974; Didday, 1976). Computer simulations were conducted to analyze how interactions among retina, optic tectum and pretectum may give frogs and toads the ability to discriminate between prey and predator stimuli. Results from simulations have allowed us to reproduce empirical observations, to suggest new experiments, and to postulate what neural mechanisms might be involved in some phenomena related to prey-catching orienting behavior, with direction invariance of prey-predator recognition being a consequence of tectal architecture, and size preference and response latency depending on the motivational state of the animal.     

The protective effect of dietary fish oil on murine lupus

Dietary marine lipids markedly reduce the severity of glomerulonephritis and its associated mortality in inbred strains of mice developing autoimmune disease, a model for human systemic lupus erythematosus. We report here the influence of varying the dose of menhaden oil and the timing of its administration on the mortality of female (NZB x NZW) F₁ mice. After ingesting 25 wt% menhaden oil (MO) for periods of 1.5 weeks to 12 months, there was a stable content of tissue n-3 fatty acids, with total n-3 fatty acids of 28% and 35% in spleen and liver, respectively. The extent of protection from mortality was dependent on the dose of MO with marked protection at doses of 11 to 25%, marginal protection at 5.5% and no protection at 2.5% MO. Delay in the institution of MO until ages 5 or 7 months still resulted in large reductions of mortality. Conversely, institution of a MO diet from 6 weeks until ages 5 to 7 months followed by a change to beef tallow resulted in little protection. Serum levels of 4 cyclooxygenase products were reduced ranging from 26 to 76% in mice fed MO diets, compared to mice fed beef tallow, based on radioimmunoassay. The degree of reduction of mortality on different doses of MO was correlated best with tissue levels of C22:5, and levels of C20:5 and C22:6 were similar at high and low doses of MO, suggesting that levels of 22:5 may be related to the protective effects of marine lipids on autoimmune disease.     

Dexamethasone-resistant cystic fibrosis fibroblasts show cross-resistance to sex steroids

Diploid skin fibroblasts derived from individuals with the autosomal recessive disease, cystic fibrosis (CF), were shown previously to be significantly more resistant to the cytotoxicity of dexamethasone, a glucocorticoid hormone, than were normal human fibroblasts. Here cystic fibrosis fibroblasts are also shown to be more resistant than normal human fibroblasts to the cytotoxic effects of the sex hormones, 17 beta-estradiol, dihydrotestosterone and progesterone. Since cells are believed to contain different receptors for each of the steroid hormones, it is not probable than the resistance of CF cells to these hormones results from a receptor deficiency. This was shown by the fact that CF cells were found to exhibit the same receptor activity as normal cells for 3-H-dexamethasone. Furthermore, neither normal human nor CF fibroblasts could be demonstrated to contain detectable receptor activity for 3H-17 beta-estradiol. In addition, the studies of fibroblast killing by hormones led to the further interesting observation that normal human diploid fibroblasts, regardless of the sex of the tissue donor, are sensitive to killing by each of the sex hormones. These findings suggest that the cytotoxic effects of the steroid hormones may be observed independently of the specific hormone receptors. The studies reported here thus suggest that the resistance of CF cells to the different steroid hormones is probably the result of a defect in a pathway in cellular steroid hormone metabolism other than that involving receptors.     

Use of cryostat sections for measurement of Ca2+ uptake by sarcoplasmic reticulum.

Calcium uptake of cardiac muscle and fast-twitch and slow-twitch skeletal muscle of rabbit was measured in 10-μm thin sections. Ca²⁺ uptake showed K⁺, Mg²⁺, Ca²⁺, and oxalate dependence and required ATP. The contribution of mitochondria to the Ca²⁺ uptake could be ruled out, since inclusion of ruthenium red or sodium azide in the medium did not show inhibition. The method, which avoids unphysiological fragmentation of the sarcoplasmic reticulum, has the added advantage of requiring only 0.1 to 0.3 g of muscle and permitting simultaneous histochemical studies from the same muscle block.     

Cell-free translation analysis of messenger RNA in echinoderm and amphibian early development

A wheat germ cell-free translation system has been used to analyze populations of abundant messenger RNA from sea urchin eggs and embryos and from amphibian oocytes and ovaries. We show directly that sea urchin eggs and embryos contain translatable mRNA of three general classes: poly(A)⁺ mRNA, poly(A)^? histone mRNA, and poly(A)^? nonhistone mRNA. Additionally, some histone synthesis appears to be promoted by poly(A)⁺ RNA. Sea urchin eggs seem to contain a higher proportion of prevalent poly(A)^? nonhistone mRNAS than do embryos. Some differences in the proteins encoded by poly(A)⁺ and poly(A)^? RNAs are detectable. Many coding sequences in the egg appear to be represented in both poly(A)⁺ and poly(A)^? RNAs, since the translation products of the two RNA classes exhibit many common bands when run on one-dimensional polyacrylamide gels. However, some of this overlap is probably due to fortuitous comigration of nonidentical proteins. Distinct stage-specific changes in the spectra of prevalent translatable mRNAs of all three classes occur, although many mRNAs are detectable throughout early development. Particularly striking is the presence of an egg poly(A)^? mRNA, encoding a 70,000–80,000 molecular weight protein, which is not detected in morula or later-stage embryos. In amphibian (Xenopus laevis and Triturus viridescens) ovary RNA, the translation assay detects the following three mRNA classes: poly(A)⁺ nonhistone mRNA, poly(A)^? histone mRNA, and poly(A)⁺ histone mRNA. Amphibian ovary RNA appearently lacks an abundant poly(A)^? nonhistone mRNA component of the magnitude detectable in sea urchin eggs. mRNA encoding histone-like proteins is found in the very earliest (small stage 1) oocytes of Xenopus as well as in later stage oocytes. During oogenesis there appear to be no striking qualitative changes in the spectra of prevalent translatable mRNAs which are detected by the cell-free translation assay.     

Interpretation of the low-angle meridional neutron diffraction patterns from collagen fibres in terms of the amino acid sequence

Measurement of fully corrected, low angle meridional neutron diffraction intensities from native collagen fibres, in a full range of H₂O/D₂O contrasts, is described. The observed contrast dependence of the intensities of the first 12 orders of 670 A (D) axial periodicity is fitted to a general quadratic theory of contrast variation. The observed first order contrast dependence is compared to predictions based on the amino acid sequence, assuming different extents of chemical H/D exchange, and found to be consistent with complete non-carbon linked H/D exchange except for 1 to 1.6 hydrogen atoms per gly-X-Ytriplet involved in H-bonding. Both X-ray and neutron diffraction data in a variety of contrasts are consistent with a unique model for the axially projected structure of native collagen fibrils based on the amino acid sequence. This model if characterized by average axial residua translations in the NH₂ and COOH terminal, non-triple helical telopeptides, expressed as multiples of the triple helical residue translation, of 0.85 ± 0.05 and 0.7 ± 01 respectively, with D = 235 ± 1 residues.     

Mechanical Land Clearing to Promote Establishment of Coastal Sandplain Grassland and Shrubland Communities

The decline in grasslands and other species‐rich early successional habitats on the coastal sandplains of the northeastern United States has spurred management to increase the area of these declining plant communities. We mechanically removed overstory oak and applied seed from a nearby sandplain grassland on the island of Martha’s Vineyard, Massachusetts, to evaluate this technique for creating an open oak community able to support sandplain herbaceous species. We compared vegetation structure and composition before and after clearing in an area of total tree removal (clearcutting), an area where 85% of tree basal area was removed (savanna cutting), and in adjacent coastal oak forest. Plant responses to clearcutting and savanna cutting were similar. Sandplain herbs colonized at high frequencies after seeding and increased herbaceous cover from less than 7% before clearing to 22–38% three growing seasons later. Pennsylvania sedge (Carex pensylvanica) increased in cover approximately 6‐fold, accounting for 84–90% of the increased herbaceous cover. Other native ruderals and exotic herbs reached 2 and less than or equal to 1%, cover, respectively, after three years. Species richness across cleared treatments increased from 30 to 79 species. All forest species were retained. Forest shrubs and trees initially declined from their dominant cover but rebounded after three years. Tree clearing plus seeding appeared to be a viable management practice for increasing cover of herbaceous sandplain species while causing minimal increases in exotic herbaceous cover. The long‐term persistence of sandplain herbs may require periodic disturbances that limit woody regrowth.