Studies of enzyme polymorphisms in the Kamuela population of Drosophila mercatorum. II. Evaluation of glycolytic intermediates

A simple and effective cryogenic procedure for the extraction of glycolytic intermediates from whole Drosophila has been developed. This procedure gives consistent results when a measure (µM/liter/OD₂₆₀) is adopted which corrects for differences in extraction efficiency. Using this measure and a homozygous strain of D. mercatorum, there are no significant differences among extracts for the levels of any of the 15 glycolytic intermediate or energy molecules considered. The profile of means is consistent across experimental designs and instrument types. Coefficients of variation are well below 50% for most variables. The methodology presented has the statistical power to detect a mean change of 10 to 50% using an experimental design which requires as few as 32 observations. The estimated energy charge for resting Drosophila from these studies is the expected value of 0.86.     

Trichomonas gallinae in columbiform birds from the Galapagos Islands

Domestic pigeons were introduced into the Galapagos Islands in 1972 or 1973. There is a high prevalence of Trichomonas gallinae among them and some evidence of canker. Trichomonas gallinae can be found also in endemic Galapagos doves in the vicinity of Puerto Ayora on Santa Cruz Island. Doves examined on pigeon-free islands were not found infected.     

The apparent absence of serotonin-mediated vascular thermogenesis in perfused rat hindlimb may result from vascular shunting

Vasoconstriction by norepinephrine, angiotensin II and vasopressin in the constant-flow perfused rat hindlimb is associated with increased oxygen uptake and has given rise to the concept of vascular thermogenesis. In the present study serotonin (5-hydroxytryptamine, 5HT) was found to inhibit oxygen uptake by up to 40% in a dose dependent manner whilst inducing vasoconstriction in this model, whereas norepinephrine increased oxygen consumption by up to 100% during vasoconstriction. This contrasted with the perfused isolated rat mesenteric artery arcade in which serotonin stimulated oxygen uptake by up to 130% in association with vasoconstriction in a dose dependent manner similar to the previously described norepinephrine induced vascular thermogenesis in this arterial preparation. In both perfusion systems, changes in pressure and oxygen uptake mediated by serotonin were completely blocked by ketanserin. These results and evidence from dye washout studies suggest that serotonin-mediated vascular thermogenesis, if it occurs in the constant-flow hindlimb, is masked by vascular shunting.     

The CD20 (Bp35) antigen is involved in activation of B cells from the G0 to the G1 phase of the cell cycle

Two monoclonal antibodies, 1F5 and B1, directed against the CD20 (Bp35) antigen were found to have both stimulatory and inhibitory effects on B cells. 1F5, but not B1, induces small resting tonsillar B cells and prolymphocytic leukemia cells to enlarge, to rapidly increase their RNA synthesis, and to become responsive to growth factors present in mixed lymphocyte reaction supernatants. In addition, 1F5 induces a moderate increase in thymidine uptake, which is accompanied by enhanced viability of the cells, but not by any increase in total cell number or by any detectable entry into S phase or mitosis. Taken together, these observations suggest that 1F5 can initiate transition from the G0 to the G1 phase of the cell cycle. The fact that all the changes observed can be inhibited by low concentrations (I50 = 50 ng/ml) of cyclosporin A is further evidence that 1F5 is involved at an early stage of B cell activation. Because both 1F5 and B1 belong to the IgG2a subclass, differences in their activities are likely to reflect their different epitope specificities. Although only 1F5 had stimulatory activity, both 1F5 and B1 strongly inhibited B cell differentiation to immunoglobulin secretion. Possible explanations for the dual activities of 1F5 and implications for the role of the CD20 antigen in B cell differentiation are discussed.     

Mg2+ inhibition of Na2+-stimulated Ca2+ release from brain mitochondria

Magnesium has been shown to modulate the Na+-stimulated release of Ca2+ (Na/Ca exchange) from brain mitochondria. The presence of 5 mM MgCl2 extramitochondrially inhibits the Na/Ca exchange as much as 70%. Additionally, Na+-stimulated Ca2+ release is enhanced by the presence of divalent chelators, this stimulation also being inhibited by the addition of excess Mg2+. The inhibitory effect of Mg2+ and the enhancement by chelating agents were both reversible. Heart mitochondria exhibit a similar enhancement of Na/Ca exchange by chelators and inhibition by MgCl2, though not as pronounced.     

Systematic implicationsofflavonoid patterns in Haplopappus segregates

Complex flavonoid patterns are correlated with primitive woody taxa and high base chromosome numbers among Haplopappus segregates. Variability among flavonoid complements supports the polyphyletic interpretation of the genus.     

Variability of Temperature, pH, and Moisture in an Aerobic Composting Process

This study measured the environmental variability which exists in a commercial aerobic composting process. The specific process studied is carried out in six decomposition cells which present six different phases of the process. Temperature, pH, and moisture content were determined in several randomly chosen sample sites in each cell, both at the beginning and at the end of the time the material was left in the cell. Temperature and pH varied greatly from one sample site to another in each cell, whereas moisture content was less varied. A significant rise in both temperature and pH was observed at two stages of degradation.     

Oxidant membrane injury by the neutrophil myeloperoxidase system. II. Injury by stimulated neutrophils and protection by lipid-soluble antioxidants

The stimulated human neutrophil can damage a variety of target cells, and in some models, a mechanism involving secretion of myeloperoxidase and H2O2 has been demonstrated. We explored the characteristics of this cell-cell interaction by using neutrophils and our recently described liposome model target cell system. Exposure of 51Cr-labeled liposomes to phorbol myristate acetate-stimulated human neutrophils resulted in release of 25 to 30% of the radioactivity. 51Cr release was abrogated by omission of the neutrophils, the phorbol ester or halide (iodide), replacement of the phorbol by an inactive congener, or addition of azide, cyanide, or catalase. Neutrophils from patients with hereditary absence of myeloperoxidase (MPO) or a failure of H2O2 formation (chronic granulomatous disease) did not cause liposome lysis unless purified MPO or a source of H2O2, respectively, was added. These data indicate that 51Cr release from liposomes is a consequence of the secretion of MPO and H2O2, which combine with extracellular halides to form a membrane lytic system. The influence of liposome composition on injury was then examined, with a focus on physiologically relevant lipid soluble antioxidants. Liposomes containing either alpha-tocopherol (0.33 to 1.67% of molar fraction of lipid) or beta-carotene (1.67% of molar fraction of lipid) were markedly resistant to lysis by the cellfree MPO-H2O2-chloride system. When the major structural lipid phosphatidyl choline was replaced by dipalmitoyl phosphatidyl choline, a synthetic phospholipid with no oxidizable double bonds, the resultant liposomes were totally resistant to lysis by the MPO-H2O2-chloride system. The addition of iodide to this system (i.e., both chloride and iodide present) changed the pattern of protection dramatically in that alpha-tocopherol and beta-carotene were no longer protective and the resistance of dipalmitoyl phosphatidyl choline liposomes was partial rather than complete. In contrast to iodide, the addition of bromide or thiocyanate did not have a major effect on the protection by antioxidants. Finally, we demonstrated protection by alpha-tocopherol or dipalmitoyl phosphatidyl choline against liposome lysis by phorbol-activated neutrophils. These studies illustrate the use of model phospholipid membranes in the characterization of oxygen-dependent cell-mediated cytotoxicity. Activated neutrophils lyse liposome targets through a MPO-dependent mechanism. Target properties, especially the content of lipid-soluble antioxidants, have a marked influence on susceptibility to lysis.(ABSTRACT TRUNCATED AT 400 WORDS)     

Aminoacyl-tRNA synthetase mutants degrade protein at a normal rate.

The stability of both rapidly and slowly degraded proteins in wild type CHO cells is similar to that in three ts aminoacyl-tRNA synthetase mutants at both permissive and non-permissive temperatures, although the degree of tRNA charging in the synthetase mutants differs considerably with temperature. These results indicate that the altered rate of protein breakdown seen under a variety of physiological conditions in eukaryotic systems is not mediated by uncharged tRNA.