Mechanism of Agonist-Induced Down-Regulation and Subsequent Recovery of Muscarinic Acetylcholine Receptors in a Clonal Neuroblastoma X Glioma Hybrid Cell Line

The mechanisms of carbachol-induced muscarinic acetylcholine receptor (mAChR) down-regulation, and recovery following carbachol withdrawal, were studied in the neuroblastoma x glioma hybrid NG108-15 cell line by specific ligand binding assays. N-[3H]Methylscopolamine ([3H]NMS) and [3H]quinuclidinyl benzilate ([3H]QNB) were used as the ligands for the cell surface and total cellular mAChRs, respectively. Exposure of cells to 1 mM carbachol for 16 h decreased the specific binding of [3H]NMS and [3H]QNB by approximately 80%. Bacitracin (1-4 mg/ml) and methylamine (1-15 mM), inhibitors of transglutaminase and of endocytosis, prevented agonist-induced loss of surface mAChRs. Pretreatment of cells with the antimicrotubular agents nocodazole (0.1-10 microM) and colchicine (1-10 microM) prevented carbachol-induced loss of [3H]QNB binding, but not that of [3H]NMS binding. These results indicate that agonist-induced mAChR down-regulation occurs by endocytosis, followed by microtubular transport of receptors to their intracellular degradation sites. When carbachol was withdrawn from the culture medium following treatment of cells for 16 h, receptors recovered and were incorporated to the surface membrane. This recovery process was antagonized by monovalent ionophores monensin (0.1 microM) and nigericin (40 nM), which interfere with Golgi complex function. Receptor recovery was also prevented by the antimicrotubular agent nocodazole. Thus, recovery of receptors appears to be mediated via Golgi complex and microtubular transport to the surface membrane.     

Seed Banks and Species Richness Potential of Coal Slurry Ponds Reclaimed as Wetlands

Seed bank experiments are described to assess the species richness potential of coal slurry ponds reclaimed as wetlands (ranging from 6 to more than 40 years old). Experimental treatments test the drawdown and flooded conditions characteristic of the vegetation dynamics of emergent wetlands in the Upper Mississippi Valley. More seedlings, primarily annuals, emerged from exposed wet sediments (freely drained) than under continuous flooded sediments in cold ponds (339 versus 136 seedlings m^?2, respectively) and in natural ponds (163 versus 47, respectively). More seeds were produced by plants established in freely drained conditions than under flooded conditions from sediments in the coal ponds (26546 versus 1842 seeds m^?2, respectively) and the natural ponds (28430 versus 4526, respectively). Similarly, more biomass was also produced by these plants in freely drained than under flooded conditions in coal ponds (118 versus 47 g m^?2, respectively) and natural ponds (118 versus 52, respectively). Fertilization (NPK) did not affect germination for the most part, but it did affect seed set and biomass production, especially for C₄ annuals such as Echinochloa crusgalli and Panicum dichotomiflorum. I propose that lime (calcium carbonate) and fertilizer be applied during the first few scheduled drawdowns for these coal slurry ponds reclaimed as wetlands to increase the number of species and to allow their more rapid development as self-sustaining systems.     

Neo-tropical felid activity patterns in relation to potential prey and intraguild competitors in the Calakmul Biosphere Reserve,Mexico

Predator behaviors influence, and are influenced by, prey and competitor behaviors. Jaguars (Panthera onca), pumas (Puma concolor), and ocelots (Leopardus pardalis) coexist throughout their geographic range as the three largest predators in a multi-predator community across diverse environments. This study tested for non-random segregation and overlap in the activity patterns of these felids and their shared prey in the southern buffer zone of the Calakmul Biosphere Reserve, in southern Mexico, using camera traps during February to August 2019. We detected little temporal segregation between the nocturnal activities of jaguars, pumas, and ocelots, although pumas were more active closer to dawn. Jaguars had low activity overlap with species likely to be common prey, whereas ocelots had high overlap with their potential prey. Pumas displayed finer-scale similarities in activity with species likely to be common prey. In an understudied area of conservation importance, this study shows that temporal segregation is an unlikely mechanism of coexistence. Further research should incorporate spatio-temporal avoidance and dietary differences to improve our understanding of the mechanisms that drive coexistence between generalist species in a diverse assemblage of threatened felids. Abstract in Spanish is available with online material.     

Prostaglandin E and mitogenic stimulation of human lymphocytes in serum-free medium

Sera used in cell cultures contain significant amount of prostaglandins (PGs). In order to vaoid any effects of contaminating PGs, the present study employed a serum-free culture medium and confirmed the inhibitory effect of prostaglandin E (PGE) on the human lymphocyte activation which had been observed previously employing a serum-containing medium. PGE₁ displayed a significantly stronger inhibitory effect on the cells than previously shown. Furthermore, reported enhancement of PGE synthesis by mitogen-activated lymphocytes could not be reproduced.     

The effective configuration of a molecule in ultrafiltration: Its definition and determination

Summary The time taken to descend through a measured depth of glycerol was recorded for some metal boxes, cylinders, ellipsoids and spheres having axial ratios from 0.2 to 2.4. The results were used to determine the relation between each nonspherical form, its axial ratio, and the radius of its dynamically equivalent sphere with respect to glycerol. Expressed in dimensionless terms, these relations closely predict thereal gross configuration of four regularly shaped molecules (inulin, ribonuclease, hemoglobin and hen egg-white lysozyme), and sensibly predict theeffective gross configuration of one irregularly shaped molecule (Gl immunoglobulin) from their known axial ratios and Stokes-Einstein radii. Similarly, they predict theeffective gross configuration of a molecule whose real shape is not known (serum albumin). Since the relations established for the models apply to all the molecules considered, it seems likely that they can be used in conjunction with membrane-diffusion studies (E. Middleton, 1975,J. Membrane Biol. 20:347) and free-diffusion studies to determine the effective configuration in ultrafiltration of any molecule.     

Inhibition by quercetin of thyroid hormone stimulation in vitro of human red blood cell Ca2+-ATPase activity

Human red blood cell membrane Ca2+-ATPase activity is stimulated in vitro by physiological concentrations of thyroid hormone. Quercetin, a flavonoid that inhibits several membrane-linked ATPases, suppressed thyroid hormone action on red cell Ca2+-ATPase activity and also interfered with binding of the hormone by red cell membranes. These effects of quercetin were dose-dependent over a range of concentrations (1-50 microM). In contrast, in the absence of thyroid hormone, quercetin at low concentrations stimulated Ca2+-ATPase activity and at 50 microM inhibited the enzyme. The effects of quercetin at low concentrations (1-10 microM), namely, stimulation of Ca2+-ATPase and inhibition of membrane-binding of thyroid hormone, mimic those of thyroid hormone and are consistent with the thyronine-like structure of quercetin. At high concentrations, quercetin is generally inhibitory of Ca2+-ATPase activity. Chalcone, fisetin, hesperetin and tangeretin are other flavonoids shown to reduce susceptibility of membrane Ca2+-ATPase to hormonal stimulation.