Search PubMed⌕ Search

Biomedical subjects

D Mackay

Publications and source records attributed to D Mackay.

At least 55 records · Page 3Linked to original sources
PubMed1997

Methylmercury decreases IL-1beta immunoreactivity in the nervous system of the developing frog Xenopus laevis.

In aquatic ecosystems, mercury can become methylated and act as a potent environmental toxin, producing developmental and neurotoxic effects in a variety of species, including frogs. Molecular indicators provide a means of assessing exposure to methylmercury and for understanding how it and other environmental toxins alter cellular function. Molecules such as growth or survival factors, and cytokines are good candidates for molecular indicators of exposure and/or damage because they are intimately related to cell and molecular processes that underlie normal growth and function. The cytokine, IL-1beta, was measured in whole frog embryos using Western blot methods and in specific structures using immunocytochemistry after exposure to 0, 10, 25, 50 and 100 parts per billion (ppb) methylmercury chloride (mmc). We observed no significant changes in total IL-1beta in whole embryo extracts. However, statistically significant decreases in IL-1beta were observed in the Vth cranial ganglion and myotomal blocks of Xenopus laevis embryos exposed to concentrations greater than or equal to 50 ppb mmc. In addition, increased mortality and alterations in gross morphology and behavior were altered by these same concentrations of mmc. Thus, frog embryos are highly susceptible to low levels of mmc contamination, and IL-1beta is an indicator of mmc exposure in the nervous system.

Animals↗
PubMed1994

A new method for estimating dissociation constants of competitive and non-competitive antagonists with no prior knowledge of agonist concentrations.

1. A method is presented which enables the dissociation constant (KI) of a competitive, pseudo-irreversible or non-competitive antagonist-receptor complex to be estimated without knowledge of agonist concentrations. 2. The technique has been tested using sets of concentration-response data which simulated these various types of antagonism. 3. The points for each set of simulated data could be plotted both as agonist concentration-response curves at fixed antagonist concentrations and vice versa, producing paired data sets. 4. pKI-values were estimated from such paired data sets using appropriate graphical and computer curve-fitting methods. 5. For competitive antagonism, for each paired data set the computer curve-fitting techniques gave the same value for pKI, assuming drug-receptor interaction to be 1:1 and agonist concentrations to be known. 6. When agonist concentrations were assumed unknown, pKIS could not be estimated by the conventional method (using agonist dose-ratios) but could still be obtained (for competitive, pseudo-irreversible and non-competitive antagonism) by the new method. 7. This new method should be especially useful for measuring dissociation constants of antagonists against neuronally- or ionophoretically-released agonists. It may also be useful when agonist is applied exogenously, especially if suitable drugs are not available to block agonist uptake and/or metabolism.

Binding Sites↗
PubMed1994

pKI values of prazosin and idazoxan for receptors stimulated by neuronally released transmitter in the epididymal portion of rat isolated vas deferens.

1. A new method has been used to measure pKI values of prazosin and idazoxan against neuronally-released transmitter in the epididymal portion of the rat isolated vas deferens. The most reproducible results were obtained with a prolonged antagonist equilibration time (1 h). 2. Under these conditions the pKI of prazosin was practically unaffected by addition of alpha, beta-methylene-adenosine-5'-triphosphate (10 microM) to desensitize purinoceptors. Addition of desmethylimipramine (DMI) (0.3 microM) produced a small, but statistically non-significant, reduction. 3. The same method has been used to measure the pKI of prazosin against exogenous noradrenaline. In the latter case addition of DMI (0.3 microM) and corticosterone (30 microM) together produced a statistically significant reduction in the apparent pKI of prazosin. 4. The new method for estimating pKI values shows that DMI itself acts either pseudo-irreversibly or non-competitively and may be reducing the apparent pKI of prazosin. 5. The pKI values obtained for prazosin and idazoxan against neuronally-released transmitter are in good agreement with those obtained by other workers for the actions of these drugs on alpha-adrenoceptors.

Adenosine Triphosphate↗
PubMed1993

Cyclin A-mediated inhibition of intra-Golgi transport requires p34cdc2.

An in vitro assay was used to study the role of p34cdc2 in cyclin A-mediated vesicular transport inhibition. It was shown that the S-phase kinase p33cdk2 reduced the effect of cyclin A on transport assays performed with sHeLa cytosol, even though histone kinase was strongly activated. Also, transport with FT210 cytosol (which is temperature-sensitive for p34cdc2) was inhibited by cyclin A only at the permissive temperature. However, the phosphatase inhibitor microcystin inhibited transport without any requirement for p34cdc2 activity. These results show that transport is inhibited by cyclin A via p34cdc2, and also by another kinase, possibly downstream of p34cdc2.

Alkaloids↗
PubMed1993

Inhibition of intra-Golgi transport in vitro by mitotic kinase.

It has previously been shown that exocytic and endocytic membrane traffic are inhibited in mitotic mammalian cells. Here we have used a cell-free intra-Golgi transport assay supplemented with heterologous cytosols to mimic this effect in vitro. Cytosols with high histone kinase activity, made either from mitotic cells or by cyclin A treatment of interphase cells, inhibited intra-Golgi transport by up to 75%. Inhibition of transport was reversed by the kinase inhibitor staurosporine or by reduction in ATP levels leading to inactivation of histone kinase. The data indicate that cell cycle control of intra-Golgi transport is due to a reversible modification of cytosol, and this assay system may be used to study the molecular mechanism of mitotic transport inhibition in mammalian cells.

Alkaloids↗
PubMed1993

Residue-based interpretation of toxicity and bioconcentration QSARs from aquatic bioassays: polar narcotic organics.

Bioconcentration and toxicity estimation for a group of substituted phenols often categorized as "polar narcotics" can be confounded by pH-dependent ionization. Two methods of correction for ionization were applied to toxicity data obtained by U.S. EPA-Duluth for fathead minnows exposed to 30 different phenols in 37 bioassays. Toxicity QSARs with corrected data were substantially different from those obtained with raw toxicity data. When ionization-corrected toxicity data were used in the critical body residue (CBR) estimation process previously successful with neutral narcotic organics, several categories of CBR, apparently related to different modes of toxic action, resulted. Published data on lethal CBR for substituted phenols were in general agreement, although such information is limited. Elimination half-life rate constants, estimated from nonlinear curve fitting to time-toxicity information, were relatively constant for the Duluth bioassay data, averaging 0.3 days. Half-life information for small aquatic organisms, both from toxicity- and bioconcentration-based tests in the literature, was in a similar range. Much of the relatively high variability encountered experimental data for substituted phenols may in large part be due to differences in metabolic degradation between chemicals and species.

Animals↗
PubMed1991

Analysis of the binding of fluorescent ligands to soluble proteins. Use of simultaneous non-linear least squares regression to obtain estimates of binding parameters.

The binding of three fluorescent ligands (warfarin, dansylsarcosine and 1-anilino-8-naphthalene sulphonate) to human albumin was analysed using simultaneous non-linear least squares regression analysis. Both mock and actual fluorescence data were examined and the results indicated that reliable estimates of the binding parameters as well as the molar fluorescence of bound ligand could be obtained. The advantage of this method of analysis is that it makes full use of all the experimental data and it eliminates the need for the graphical procedures usually employed to estimate the molar fluorescence of bound ligand and its binding constants. This type of analysis can be extended to other systems where some physical property of the bound ligand varies with increasing protein concentration.

Albumins↗
PubMed1991

The membrane spanning domain of beta-1,4-galactosyltransferase specifies trans Golgi localization.

Chimeric cDNAs were constructed so as to generate hybrid proteins in which different parts of the N-terminal domain of the human invariant chain were replaced by equivalent sequences from the trans Golgi resident enzyme, beta-1,4-galactosyltransferase. The cytoplasmic and membrane spanning domains of galactosyltransferase were found to be sufficient to retain all of the hybrid invariant chain in trans Golgi cisternae as judged by indirect immunofluorescence, treatment with brefeldin A and immuno-electron microscopy. As few as ten amino acids corresponding to the lumenal half of the membrane spanning domain of the Golgi enzyme sufficed to localize most of the hybrid invariant chain to the trans cisternae. A cytoplasmic domain was necessary for complete retention as assessed by flow cytofluorometry but could be provided either by galactosyltransferase or by invariant chain. This suggests that the cytoplasmic domain plays a role accessory to the membrane spanning domain, the latter mediating compartmental specificity.

Amino Acid Sequence↗
PubMed1991

Interpreting aquatic toxicity QSARs: the significance of toxicant body residues at the pharmacologic endpoint.

Aquatic QSAR investigations employ bioassay data where a biological response is associated with a toxicant concentration in the exposure water. Although convenient this ignores the fundamental principle that the response is more closely related to an amount present in the organism. This problem has been examined using acute toxicity and bioconcentration QSARs employing log Kow as the molecular descriptor. Combining these QSAR relationships enables estimates of the amount of toxicant in organisms to be made. For acute narcosis (50% mortality) the toxicant level in the hydrophobic (lipid) phase of an exposed organism is, as a first approximation, constant at approximately 50 mmol l-1 of lipid. For the acute toxicity of a variety of hydrophobic narcotic organics examined by US EPA-Duluth in tests with fathead minnows, this corresponds to a whole-body residue of about 4 mmol kg-1. Although the level in the hydrophobic phase remains constant, for hydrophilic chemicals (log Kow less than 1.5) the contribution of the hydrophilic (water) phase of the organism dominates and total body residues should be similar to the respective threshold LC50s. This has important implications in simple pharmacokinetic modelling and in interpreting residue levels in organisms collected in environmental monitoring studies.

Animals↗
PubMed1990

Interpretation of relative potencies, relative efficacies and apparent affinity constants of agonist drugs estimated from concentration-response curves.

Differences in the relative potencies of agonists have been used successfully in the past to classify receptors. Such use of agonists can be justified on the basis of ideas and equations developed using the occupancy model of drug action. However the occupancy model makes no allowance for possible complications which may arise when the drug-receptor complex interacts with a transducer-effector system. For some receptor-effector systems use of an equilibrium ternary complex model may be better than use of the occupancy model but the former still does not take into account the possible effect of guanosine-5'-triphosphate on the system. A steady-state version of the ternary complex model has therefore been analysed to explore possible interpretations of relative potencies, relative efficacies and apparent affinity constants estimated from concentration-response curves. It is concluded that for agonists which act on receptors which function through G-proteins these pharmacological parameters may depend on the concentration of the relevant G-protein in the cell membranes and on the intracellular concentrations of guanosine-5'-triphosphate and guanosine-5'-diphosphate. If these concentrations vary appreciably between tissues then the parameters are also likely to vary, even for a single receptor-transducer system. It follows that the use of such agonist parameters to classify receptors or receptor-transducer systems is not likely to be totally dependable. It is also possible that agonists which interact with only one receptor-transducer system may show selectivity between tissues with different concentrations of G-proteins and of guanine nucleotides.

Animals↗
PubMed1990

Solubilities of pesticide chemicals in water. Part I: Environmental physical chemistry.

It is hoped that this review of the equilibrium aqueous solution thermodynamics and environmental partitioning tendencies of chemicals will be valuable in elucidating the behavior of pesticide chemicals in the environment and in promoting the development of more reliable correlations between physical chemical measurements and environmental partitioning coefficients. Ultimately the success of both the thermodynamic and environmental interpretations depend on having reliable, critically reviewed data from both laboratory and the "field."

Chemical Phenomena↗
PubMed1990

Agonist potency and apparent affinity: interpretation using classical and steady-state ternary-complex models.

The relative potencies of agonists have been used in the past to classify receptors. This could be justified on the basis of ideas and equations developed using the occupancy model of drug action. Although attempts have been made recently to develop methods of analysis based on other models, these have not been entirely satisfactory. In this article Dennis Mackay explores possible interpretations of relative potency, apparent affinity and efficacy on the basis of a steady-state ternary-complex model. He concludes that use of these quantities to classify receptor-transducer systems may not be entirely dependable. However the steady-state model can explain why use of agonists to classify receptors has been successful in the past and why properties of agonists deduced from ligand binding studies may differ from their properties deduced from experiments on intact tissues.

Animals↗
PubMed1989

Correlation of tissue, blood, and air partition coefficients of volatile organic chemicals.

The physical chemical factors controlling partition coefficients between air, water, blood, and various tissues are discussed. It is suggested that improved insights into the relations between partition coefficients, which are frequently expressed as correlations, may be obtained by viewing the partition coefficients as ratios of solubilities or pseudosolubilities. A simple, novel correlation approach is developed and applied to 24 volatile organic chemicals, which enables tissue/blood, tissue/air, and blood/air partition coefficients to be estimated from water solubility and vapour pressure. An illustration is presented in which these solubilities are used to calculate the equilibrium distribution of dichloromethane between air, blood, and various tissues.

Air↗