Transport and control of Ca by pigeon erythrocytes. II. Evidence against a simple feedback control of cell Ca and evidence for the involvement of more than one pool

Cells were subjected to a range of ⁴⁵Ca²⁺ influx loads with A23187. We measured cell ⁴⁵Ca²⁺ with time and A23187 dose, and the apparent ⁴⁵Ca²⁺ influxes (≡“J(in,app)”) at Ca²⁺ steady state. We also measured endogeneous exchangeable and total cell Ca²⁺, which were 50 and 17–220 μM respectively. The effects of A23187 and Ca²⁺ on cell ATP, swelling, net Cl⁻ permeability, and cell morphology were measured. These were modest and do not affect our conclusions.J(in,app) 3 × 10⁻⁴ [A23187]^2.9·[Ca²⁺(o)]μmoles/l·min with 92–552 μM [Ca²⁺(o)] (≡ external Ca²⁺ concentration) and 0–7 μM A23187. J(in,app) was increased an order of magnitude by vanadate and is probably much less than the true influx. The least unlikely explanation found for the high [A23187] exponent, 2.9, was that most of the Ca²⁺ crossing the membrane is expelled by the pump before it can move deeper into the cell.Calcium pumping increased rapidly in response to increased influx, but the steady state cell ⁴⁵Ca²⁺ was approximately proportional to J(in,app) rather than approximately constant between 10 and 120 μmoles/l·min with 184 μM [Ca²⁺(o)]. This is not the result expected from a simple feedback mechanism.At high A23187 doses the pump appears fully activated resulting in a linear relation between cell/medium ⁴⁵Ca²⁺ and [A23187]⁻². From the plot we calculated α≡free/total exchangeable Ca²⁺ = 0.38 ± 0.08 (n = 3) and a maximum pump rate, “Pmax” = 78 μmole/l·min. Pmax is underestimated insofar as J(in,app) is less than the true influx.     

Changes in [3H]inositol-labelled phosphoinositides of pig platelets in response to thrombin

Pig platelet phosphoinositides have been labelled with [³H]inositol and then treated with thrombin in the absence of Ca²⁺. There was a loss of labelled phosphatidylinositol 4,5-bisphosphate between 30 and 60 s after the addition of thrombin but the general picture was of increased labelling over a 4-min period. Labelling of phosphatidylinositol 4-phosphate showed no period of loss but there was an early loss of phosphatidylinositol and no increased labelling during the 4-min incubation. The small amount of lysophosphatidyl[³H]inositol in the platelets was not affected by thrombin treatment. Thrombin caused loss of [¹⁴C]arachidonate-labelled phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol.     

Biospecific fractionation matrices for sequence specific endonucleases

Fractionation of several type II specific restriction endonucleases was achieved by separation on two novel biospecific matrices. The matrices are pyran, a copolymer of divinyl ether of maleic anhydride, and Cibacron Blue F3GA, a blue dye commonly used for the calibration of molecular sieves. Both compounds are insolubilized by coupling to sepharose through a cyanogen bromide linkage and in their soluble form inhibit the restriction endonucleases which we have tested. These affinity matrices can be used to obtain restriction endonucleases from crude extracts after removal of nucleic acids. They have also proven to have a high capacity when used as subsequent steps in enzyme purification. Their additional advantage is the rapid development time and reusability of columns packed with the two matrices.     

Diagnosis of pseudomembranous colitis.

Twenty-eight patients with histologically proved pseudomembranous colitis have been seen in one hospital since July 1975. All patients with the disease had received antibiotics, six for infections not requiring operations; the other 22 cases all occurred after major surgery. All the patients had diarrhoea; six patients also had fever with clinical signs of sepsis, and three had abdominal pain thought to be due to anastomotic dehiscence after colonic resection. Pseudomembranous colitis was associated with white blood counts over 15 000/mm3 in 17 patients and albumin concentrations of less than 30 g/1 in 18. Pseudomembranous colitis was an incidental finding at necropsy in two of six patients who had not had an operation. Of the 22 patients who had had major surgery, nine died from this complication; in all except two of these cases the diagnosis was made only at necropsy. If pseudomembranous colitis is suspected on clinical grounds or if there is an unexplained complication after colorectal surgery repeat sigmoidoscopy and testing for faecal toxins should be carried out to establish the diagnosis so that prompt supportive treatment can be given.     

Quinaldine sulphate, a new anaesthetic formulation for tropical marine fishes

Quinaldine sulphate is demonstrated in this study to be a convenient and safe anaesthetic for use with tropical marine fishes. Both onset of anaesthesia and recovery are significantly more rapid than with the parent compound, quinaldine.     

High-performance liquid chromatography of p-nitrophenacyl esters of selected prostaglandins on silver ion-loaded microparticulate cation-exchange resin

A silver ion-loaded microparticulate cation-exchange resin column has been used for high-performance liquid chromatographic (HPLC) separation of the p-nitrophenacyl esters of several series of closely related prostaglandins: 8-iso-PGE₂, 11-epi-PGE₂, 5-trans-PGE₂, PGE₂, PGF_1α, PGE₁, and PGF_2α, PGA₂ and PGB₂; 15 (R)-methyl-PGE₂ and 15 (S)-methyl-PGE₂; 5-trans-PGA₂ and PGA₂; and 5-trans-PGF_2α and PGF_2α. The properties of this column are compared with those of silica-gel and reversed-phase columns.     

Contributions of subterranean termites to the “economy” of Chihuahuan desert ecosystems

Summary We examined the role of subterranean termites in decomposition of cattle dung, various herbaceous plant species and wood in a Chihuahuan desert ecosystem. From July–September, termites removed dung at a rate of 0.63 g day^-1 accounting for a percent mass loss of 19.5–100%. During the autumn subterranean termites consumed more than 50% of the leaves of the shrub Larrea tridentata, the grass Erioneuron pulchellum and annual plant Lepidium lasiocarpum and Baileya multiradiata but used very little of two other annuals Eriastrum diffusum and Eriogonum trichopes. Yucca inflorescence stalks on plots with termites lost 23% of their original mass in 30 months while those on termite free soils lost 11%. Elimination of termites resulted in reduction of fluff grass, Erioneuron pulchellum biomass, thereby affecting the structure of the ecosystem.     

A reassessment of the binding of napthaleneacetic acid by membrane preparations from maize

Naphthalene-1-acetic acid (NAA) binding by membrane fractions derived from maize has been re-evaluated. Using a computer curve-fitting procedure only one major type of NAA binding, in terms of binding affinity, could be identified. Auxins, antiauxins and structurally related compounds have been tested for their competitive effect on NAA binding and the inhibitor constants for a number of these have been determined. Extracts from various plant species have been examined for their NAA binding ability, but all showed much less binding than maize leaf or coleoptile preparations. The possibility of the NAA binding by maize extracts being due to a true hormone receptor is discussed.Abbreviations BA benzoic acid - CPIB p-chlorophenoxyisobutyric acid - 2,4-D 2,4-dichlorophenoxyacetic acid - DCB 2,4-dichlorobenzoic acid - IAA indolyl-3-acetic acid - NAA napthalene-1-acetic acid - 2-NAA napthalene-2-acetic acid - NAOA napthalene-2-oxyacetic acid - PA phenylacetic acid - PU phenylurea - 2,4,5-T 2,4,5-trichlorophenoxyacetic acid - TIBA 2,3,5-triiodobenzoic acid