Tetrahymena Telomerase RNA levels increase during macronuclear development

Telomeres, the G-rich sequences found at the ends of eukaryotic chromosomes, ensure chromosome stability and prevent sequence loss from chromosome ends during DNA replication. During macronuclear development in Tetrahymena, the chromosomes fragment into pieces ranging from 20 kb to 1,500 kb. Tetrahymena telomerase, a ribonucleoprotein, adds telomeric (TTGGGG)_n repeats onto telomeres and onto the newly generated macronuclear DNA ends. We have investigated whether telomerase RNA levels increase during macronuclear development, since such an increase might be expected during chromosomal fragmentation. The steady-state level of the telomerase RNA component was used to estimate the abundance of telomerase present in mating and nonmating Tetrahymena. Northern blot analysis revealed that in vegetatively growing Tetrahymena, there were 18,000–40,000 copies of telomerase RNA per cell. In mating cultures, the levels of RNA increased 2-to 5-fold at 9–15 h, and 1.5- to 3.5-fold in starved nonmating cultures. This increase in telomerase RNA paralleled telomerase activity, which also increased slightly in mating and starved nonmating cells. © 1992 Wiley-Liss, Inc.     

6-substituted tricyclic partial ergoline compounds are selective and potent 5-hydroxytryptamine1A receptor agents

A series of 6 tricyclic partial ergoline derivatives was analyzed using radioligand binding assays. Four agents (LY 178210, LY 254089, LY 197205, and LY 197206) display high affinity (Ki less than or equal to 1.3 nM) for 5-hydroxytryptamine1A (5-HT1A) receptor binding sites labeled by [3H]8-hydroxy- 2-(di-n-propylamino) tetralin (8-OH-DPAT) and display greater than or equal to 150 fold selectivity for the 5-HT1A over the 5-HT1D receptor binding site. The most potent agent investigated, LY 178210, is essentially inactive (Ki greater than 1500 nM) at a total of 12 other neurotransmitter receptor binding sites in the brain. Using a forskolin-stimulated adenylate cyclase assay as a model of 5-HT1A receptor function, LY 178210 was found to display partial agonist activity which was blocked by 10(-5) M (-)pindolol. These data indicate that LY 178210 is a potent and selective 5-HT1A receptor partial agonist.     

Effect of drift sampler exposure time and net mesh size on invertebrate drift density in the Njoro River,Kenya

Although invertebrate drift is an important ecological process in lotic ecosystems, very little is known about it in Kenyan rivers. The primary aim of this study was to investigate the effect of driftnet mesh size and exposure duration on drift density in 2017. Drift samples were dominated by Chironomidae, Baetidae, Simuliidae, Caenidae and Culicidae. The 100 µm mesh driftnet had the highest mean invertebrate density, followed by the 250 µm and 500 µm nets. Invertebrate drift densities decreased with increased exposure time. This study demonstrates that sampler mesh size and exposure time should be taken into account when characterising invertebrate drift in streams. Future studies should consider sampling different biotopes and during different seasons.     

Synthesis of peptidoglycan and teichoic acid in Bacillus subtilis: role of the electrochemical proton gradient.

The effects of several ionophores and uncouplers on glycerol and N-acetylglucosamine incorporation by Bacillus subtilis 61360, a glycerol auxotroph, were tested at different pH values. In particular, the effect of valinomycin on the synthesis of teichoic acid and peptidoglycan was examined in more detail in both growing cells and in vitro biosynthetic systems. Valinomycin inhibited synthesis of wall teichoic acid and peptidoglycan in whole cells but not in the comparable in vitro systems. It did not inhibit formation of free lipid or lipoteichoic acid. The results were consistent with a role for the electrochemical proton gradient in maintaining full activity of cell wall synthetic enzymes in intact cells. Such an energy source would be required for a model in which rotation or reorientation of synthetic enzyme complexes is envisaged for the translocation of wall precursor molecules across the cytoplasmic membrane (Harrington and Baddiley, J. Bacteriol. 155:776-792, 1983).     

Suitability of the modified fluctuation assay for evaluating the mutagenicity of unconcentrated drinking water

Filter-sterilized, unconcentrated tap water induced mutagenic responses (p less than 0.01) in Salmonella strain TA100 in fluctuation assays, usually with dose-related increases in positive tubes. Additional experiments were performed to study possible artifacts that could lead to falsely positive results. Determinations of bacterial survival revealed that cell populations in the tubes containing tap water were larger than in the controls. Since spontaneous mutation is a function of cell generation, the increased numbers of bacteria appeared to be responsible for the higher numbers of mutants observed. Therefore, the positive responses must be regarded as artifactual. This study suggests that survival determination should be a routine part of this method, and care should be exercised in the interpretation of positive results.     

Norepinephrine causes alpha 1-adrenergic receptor-mediated decrease of phosphatidylinositol in isolated rat liver plasma membranes supplemented with cytosol

When purified rat liver plasma membranes were incubated with norepinephrine, rat liver cytosol, and Ca2+, the amount of membrane-bound phosphatidylinositol was reduced by up to 50%. The levels of other major membrane phospholipids underwent negligible change. The decrease in phosphatidylinositol levels was not observed if norepinephrine was omitted or cytosol was absent. The disappearance of phospholipid persisted when soluble Ca2+ was depleted by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. The decrease was prevented by phentolamine, benextramine, and prazosin, but not by sotalol. The results show that norepinephrine elicits a specific alpha 1-adrenergic receptor-mediated breakdown of phosphatidylinositol in isolated plasma membranes which is dependent on the presence of cytosol.     

Lymphocyte locomotion and attachment on two-dimensional surfaces and in three-dimensional matrices

The adhesion and locomotion of mouse peripheral lymph node lymphocytes on 2-D protein- coated substrata and in 3-D matrices were compared. Lymphocytes did not adhere to, or migrate on, 2-D substrata suck as serum- or fibronectin-coated glass. They did attach to and migrate in hydrated 3-D collagen lattices. When the collagen was dehydrated to form a 2-D surface, lymphocyte attachment to it was reduced. We propose that lymphocytes, which are poorly adhesive, are able to attach to and migrate in 3-D matrices by a nonadhesive mechanism such as the extension and expansion of pseudopodia through gaps in the matrix, which could provide purchase for movement in the absence of discrete intermolecular adhesions. This was supported by studies using serum-coated micropore filters, since lymphocytes attached to and migrated into filters with pore sizes large enough (3 or 8 mum) to allow pseudopod penetration but did not attach to filters made of an identical material (cellulose esters) but of narrow pore size (0.22 or 0.45 mum). Cinematographic studies of lymphocyte locomotion in collagen gels were also consistent with the above hypothesis, since lymphocytes showed a more variable morphology than is typically seen on plane surfaces, with formation of many small pseudopodia expanded to give a marked constriction between the cell and the pseudopod. These extensions often remained fixed with respect to the environment as the lymphocyte moved away from or past them. This suggests that the pseudopodia were inserted into gaps in the gel matrix and acted as anchorage points for locomotion.