Modulation of opioid receptor binding by cis and trans fatty acids.

In synaptosomal brain membranes, the addition of oleic acid (cis), elaidic acid (trans), and the cis and trans isomers of vaccenic acid, at a concentration of 0.87 mumol of lipid/mg of protein, strongly reduced the Bmax and, to a lesser degree, the binding affinity of the mu-selective opioid [3H]Tyr-D-Ala-Gly-(Me)Phe-Gly-ol ([3H]DAMGO). At comparable membrane content, the cis isomers of the fatty acids were more potent than their trans counterparts in inhibiting ligand binding and in decreasing membrane microviscosity, both at the membrane surface and in the core. However, trans-vacenic acid affected opioid receptor binding in spite of just marginally altering membrane microviscosity. If the receptors were uncoupled from guanine nucleotide regulatory protein, an altered inhibition profile was obtained: the impairment of KD by the fatty acids was enhanced and that of Bmax reduced. Receptor interaction of the delta-opioid [3H](D-Pen2,D-Pen5)enkephalin was modulated by lipids to a greater extent than that of [3H]DAMGO: saturable binding was abolished by both oleic and elaidic acids. The binding of [3H]naltrexone was less susceptible to inhibition by the fatty acids, particularly in the presence of sodium. In the absence of this cation, however, cis-vaccenic acid abolished the low-affinity binding component of [3H]naltrexone. These findings support the membrane model of opioid receptor sequestration depicting different ionic environments for the mu- and delta-binding sites. The results of this work show distinct modulation of different types and molecular states of opioid receptor by fatty acids through mechanisms involving membrane fluidity and specific interactions with membrane constituents.

Relationship between opioid-receptor occupancy and stimulation of low-Km GTPase in brain membranes.

Treatment of rat brain membranes with the irreversible opioid ligand cis-3-methylfentanylisothiocyanate (Superfit) was used to reduce gradually the number of available binding sites for the delta-selective agonist [3H][D-Ser2,Leu5]enkephalin-Thr6 ([ 3H]DSLET). Subsequently, the correlation between ligand binding and low-Km GTPase was investigated. Alkylation with 10 microM and 25 microM Superfit inactivated 66% and 71% of high-affinity (KD, 1 nM) binding sites without decreasing the affinity of the remaining sites and the stimulation of low-Km GTPase by DSLET. Following exposure of the membranes to 50 microM and 75 microM Superfit, ligand binding was confined to the low-affinity (KD, 20 nM) sites. In these membranes, the delta-agonists DSLET and [D-Pen2,D-Pen5]enkephalin still stimulated low-Km GTPase, and these effects were blocked by ICI 174864 (N,N-diallyl-Tyr-AIB-AIB-Phe-Leu-OH; AIB, alpha-aminoisobutyric acid), a delta-selective antagonist. A similar relationship between low-affinity ligand binding and GTPase stimulation was observed following alkylation of the delta-opioid receptor with the non-selective irreversible antagonist beta-chlornaltrexamine in the presence of protective concentrations of DSLET. The results reveal spare receptor sites in the coupling of the delta-opioid receptor to low-Km GTPase in brain and identify low-affinity ligand binding as a functional component in the process.

Efficient computer subroutines for interconversion of oxygen tension and saturation.

Two digital computer subroutines are described. NORSAT computes human hemoglobin saturation from oxygen tension data; NORINV performs the inverse computation. Each subroutine is based on an algorithm that is a direct mathematical inverse of the other. The effects of nonstandard conditions of temperature, pH, and CO2 tension on the saturation curve can be computed. The relative efficiency of these subroutines is compared to that of published subroutines having similar function. The new subroutines use computer time more efficiently with no loss of accuracy. The subroutines are provided in FORTRAN in a form suitable for direct inclusion in a number of user-specified programs.

Human serum immunoglobulin concentrations: prevalence of immunoglobulin deficiencies.

Classical antibody deficiency syndromes, such as sex-linked aggammaglobulinemia, are rare and relatively homogeneous in presentation. In the present investigation an unselected group of 3,213 individuals from a community health study was examined in an attempt to estimate the prevalence of the commoner and largely unclassified examples of immunoglobulin deficiencies defined by the lower 2.5 per cent of the population. The prevalence of selective IgA deficiency (an isolated absence of IgA) was 0.097 per cent and that for selective IgM deficiency was 0.03 per cent. No isolated absence of IgG was found. In addition to these deficiency syndromes, concentrations of each of the immunoglobulins were found to be highly correlated to each other.