New Approach to Information Handling in General Practice

In this paper we describe the use of punched feature cards in a general practice for 18 months. Its advantages are the low cost, speed of information retrieval, visible statistics, computer compatibility, accuracy, confidentiality, flexibility, and simplicity of setting up and collection of information. The system encourages the doctor to ask questions about his practice, and could readily be adopted in other practices.     

Receptor and G-protein-dependent regulation of turkey erythrocyte phosphoinositidase C

Several lines of experimental evidence indicate the involvement of a guanine nucleotide-dependent protein (G-protein) in the hormone-stimulated hydrolysis of phosphatidylinositol(4,5)-bisphosphate (PtdIns(4,5)P2). However, the shortcomings of available procedures for cell-free assay of hormone-stimulated phosphoinositidase C (PIC) have limited our current understanding of the molecular and mechanistic details of PIC regulation. We recently have proposed that turkey erythrocyte membranes may provide a valuable model system for studies of G-protein-dependent PtdIns(4,5)P2 hydrolysis. The membranes can be simply prepared from [3H]inositol-labelled erythrocytes and they contain a PIC activity that hydrolyses endogenous phosphoinositides and is exquisitively sensitive to guanine nucleotides. PtdIns(4,5)P2 is the principal substrate for this enzyme, there being relatively little direct hydrolysis of phosphatidylinositol 4-phosphate and no detectable hydrolysis of PtdIns. The membranes also contain a purinoceptor of the P2y subclass that is efficiently coupled to PtdIns(4,5)P2 hydrolysis both in intact cells and in the isolated membranes. 2-Methylthioadenosine trisphosphate (2-methyl-S-ATP), a specific P2y receptor agonist, has no effect upon PtdIns(4,5)P2 hydrolysis in the absence of guanine nucleotides, but greatly enhances both the potency and efficacy of PIC activation by guanine nucleotides such as GTP gamma S. GTP gamma S alone stimulates PIC activity only after a prolonged time-lag; the effect of increasing doses of 2-methyl-S-ATP is progressively to shorten this lag phase. These results suggest that the mechanism of G-protein activation involves acceleration of a nucleotide exchange reaction as has been demonstrated for the activation of adenylate cyclase in the same membrane preparation. As well as contributing valuable information on the substrate specificity of PIC and its mode of regulation by hormones, turkey erythrocytes provide a plentiful source of plasma membranes and may be useful for purification of the appropriate G-protein and PIC activities.     

Characterization of an altered membrane form of the beta-adrenergic receptor produced during agonist-induced desensitization

Incubation of 1321N1 human astrocytoma cells with 1 microM isoproterenol rapidly results in the conversion of a portion of the beta-adrenergic receptors to a membrane form that can be separated from markers for the plasma membrane by sucrose density gradient or differential centrifugation. This "light peak" form of the receptor reaches a maximal level within 10 min of incubation of cells with catecholamine. Two types of experiments suggest that the early phase of catecholamine-induced desensitization of the beta-adrenergic receptor-linked adenylate cyclase can be separated into at least two reactions. First, the agonist-induced loss of catecholamine-stimulated adenylate cyclase activity precedes the appearance of beta-adrenergic receptors in the light peak fraction by 1-2 min. Second, pretreatment of cells with concanavalin A prior to induction of desensitization blocks the formation of the light peak form of beta-adrenergic receptors without blocking the "uncoupling" reaction as measured by catecholamine-stimulated adenylate cyclase activity. Specificity for the reaction that converts beta-adrenergic receptors to the light peak form is indicated by the lack of a catecholamine-induced alteration in the sucrose density gradient distribution of muscarinic cholinergic receptors, adenylate cyclase or the guanine nucleotide-binding proteins, Ns and Ni. The light peak of beta-adrenergic receptors migrates at a density similar to that of at least a portion of the activity of galactosyltransferase, a marker for Golgi. Enzyme marker activities for lysosomes and endoplasmic reticulum are not associated with this population of beta-adrenergic receptors. Taken together, these and other data suggest that incubation of 1321N1 cells with isoproterenol results in a rapid uncoupling of beta-adrenergic receptors from adenylate cyclase which is followed by a change in the membrane form of the receptor. This latter step most likely represents internalization of receptors into a vesicular form which may then serve as the precursor state from which receptors are eventually lost from the cell.     

Results of Treatment of Thyrotoxicosis after Postoperative Relapse

Ninety patients who had a recurrence of thyrotoxicosis after thyroidectomy have been reviewed. All 10 patients who had a second operation and 18 out of 20 patients treated with a full course of antithyroid drugs relapsed. These results differ greatly from the results of treatment of the first episode of thyrotoxicosis, whether by thyroidectomy or antithyroid drugs. Radioiodine is the treatment of choice in this group of patients, despite the high incidence of hypothyroidism.     

Effects of negative air ions,noise, sex and age on maze learning in rats

An equal number of male and female rats of the King-Holtzman hybrid breed, divided into two age groups (group A₁ : 21–30 days old and group A₂ : 90–100 days old), were exposed to two levels of noise: N₀ = 30 db and N₁ = 90 db, (Ref. 0.0002μ bar), and three levels of negative air ions: I_O = no measurable ion concentration, I₁ = 7 × 10⁶ ions/cm³ and I₂ = 7 × 10⁷ ions/cm³. Time and error scores of 240 rats running a modified Lashley left-right maze with an escape-from-water motive served as criteria. A randomized complete blocks design with replications (2×3×2×2×10) was selected for treatment by analysis of variance. The results indicate that (a) the males show significantly lower error score in negatively ionized air; and (b) the females swim significantly faster than males under all investigated conditions with no apparent effect of noise or ions on their performance.

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Zusammenfassung Eine gleiche Anzahl m?nnlicher und weiblicher Ratten (n = 120) des King-Holtzman Stammes, unterteilt in 2 Altersgruppen (21–30 und 90–100 Tage) wurden zwei Ger?uschintensit?ten (30 db und 90 db) und 3 Konzentrationen negativer Luftionen I₀: nicht messbar niedrig, I₁:7×10⁶, I₂:7×10⁷ Ionen/cm³ ausgesetzt. Die Wirkung wurde geprüft an der Schwimmzeit und der Anzahl Fehler der Tiere im Lasley-Links-Rechts-Labyrinth. Das Motiv zum Schwimmen war, aus dem Wasser zu entkommen. Die Auswertung durch Varianzanalyse erfolgte unter Nachbildung randomisierter kompletter Blockentwürfe. Die Ergebnisse zeigen, dass (a) die M?nnchen bei negativ ionisierter Luft signifikant weniger Fehler machten und (b) die Weibchen unter allen Bedingungen schneller schwammen als die M?nnchen, ohne dass das L?rmniveau oder die Luftionisation darauf einen Einfluss hatten.

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Resume Un nombre égal de rats males et femelles (120 de chaque) de la race King-Holtzman futdivisé en deux groupes d'age (21 à 30 jours et 90 à 100 jours). Ils furent exposés à 2 intensités auditives (30 et 90 db) et à 3 concentrations d'ions négatifs: I₀ non mesurables (faible), I₁ : 7 × 10⁶ et I₂: 7 × 10⁷ ions/cm³. Les effets d'un tel traitement furent examinés au moyen de deux critères: le temps pris pour sortir de l'eau et le nombre de fautes commises dans un labyrinthe droitegauche de Laslay. Pour l'analyse des variances, on a adopté une répartition due au hasard avec répétitions (2×3×2×2×10). On peut en conclure que: (a) les males font moins d'erreurs dans l'air ionisé négativement et cela de fa?on significative et (b) les femelles ont nagé plus vite que les males dans toutes les conditions de l'essai. Ni le bruit ni le degré d'ionisation ne semblent avoir d'influence sur leur comportement.

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This study was supported in part by a grant from the Faculty Research Fund of the University of Oklahoma.     

Reconstitution of band 3, the erythrocyte anion exchange protein

Band 3, the erythrocyte membrane protein thought to be responsible for anion transport, was purified to near homogeneity using a Concanavalin A affinity column. Band 3 was then combined with egg lecithin, erythrocyte lipid, cholesterol, and glycophorin, the major erythrocyte sialoglycoprotein, to form vesicles capable of rapid sulfate transport. The transport activity was sensitive to prior treatment of the erythrocytes with pyridoxal phosphate-NaBH₄, a potent inhibitor of anion transport in these cells.     

Action Spectra for Conversions of Phycochrome c from Nostoc muscorum

The reversibly photochromic pigment, phycochrome c, was extracted from the blue-green alga Nostoc muscorum strain A. Action spectra were determined for in vitro conversions of the pigment from the short wavelength to the long wavelength form and vice versa. The action peak for the absorbance decrease at 650 nm is at 630 nm. During this decrease there is only a slight increase of the absorbance in the green region. Green and yellow light (maximum efficiency at 580 nm) completely restores absorbance at 650 nm. The observations are explained by the existence of three spectrally different forms of phycochrome c: P^c₆₃₀ and P^c₆₅₀ which equilibrate in darkness and P^c₅₈₀ which is reversibly photoconvertible to P^c₆₃₀. We have also measured the absorbance changes brought about by saturating irradiations with light of various wavelengths (“photostationary state spectrum”). Extreme photostationary states were obtained with about 650 nm and 500 nm light.     

Monoclonal antibodies to a nitroxide lipid hapten

The isolation and characterization of a hybridoma cell line producing a monoclonal IgG₁ antibody against a spin-label nitroxide group is described. The antibody recognizes a synthetic hapten containing linked dinitrophenyl and 2,2,6,6-tetramethylpiperidinyl 1-oxy groups, having an affinity of 3.6±1.0·10⁶ M^?1 for the soluble hapten at 25°C. The antibody binds to phospholipid vesicles containing 2 mol% of spin label-derivitized lipid (lipid hapten) with an affinity of 1.5±0.2·10⁸ M^?1. This monoclonal IgG₁ mediates the binding of hapten-bearing lipid vesicles to mouse macrophage RAW264 cells bearing Fc receptors. The cellular responses to this binding are similar to those observed previously using polyclonal rabbit anti-hapten IgG. As with the heterogeneous antibodies, the monoclonal IgG₁ is more efficient in mediating cellular uptake when the vesicles are in the ‘fluid’ physical state (dimyristoylphosphatidylcholine at 37°C) compared to ‘solid’ (dipalmitoylphosphatidylcholine at 37°C). Despite the enhanced binding of ‘fluid’ phospholipid vesicles to cells, only the ‘solid’ vesicles triggered a significant respiratory burst in RAW264 macrophages.