Intervention at an early threshold for post-hemorrhagic ventricular dilatation in preterm infants: a systematic review and meta-analysis.

Background: Very few adequately powered studies exploring early thresholds for intervention in the management of post-hemorrhagic ventricular dilatation (PHVD) in preterm infants have identified consistent neurodevelopmental advantages at 12-30 months. We aimed to conduct a meta-analysis on the efficacy and safety of early versus conservative thresholds for intervention, primarily aimed at normalizing cerebrospinal fluid (CSF) pressure, in the management of PHVD in preterm infants.

Methods: Multiple databases were searched for eligible papers, and prospective randomized trials involving preterm infants were selected.

Immune and metabolic markers for identifying and investigating severe Coronavirus disease and Sepsis in children and young people (pSeP/COVID ChYP study): protocol for a prospective cohort study.

Introduction: Early recognition and appropriate management of paediatric sepsis are known to improve outcomes. A previous system's biology investigation of the systemic immune response in neonates to sepsis identified immune and metabolic markers that showed high accuracy for detecting bacterial infection. Further gene expression markers have also been reported previously in the paediatric age group for discriminating sepsis from control cases.

[3H]benzylpempidine, a new radioligand for probing a putative channel site on nicotinic cholinergic receptors.

A study was undertaken to assess the receptor binding characteristics of [3H]4-benzylpempidine to an allosteric site on calf brain membranes associated with nicotinic cholinergic receptors and to compare the binding affinity of novel arylpempidine analogs with their ability to antagonize the behavioral effects of nicotine in mice. Scatchard analysis of the binding yielded a K(d) of 20 nM and a B(max) of 330 fmols/mg membrane protein. [3H]4-benzylpempidine appears to be a more satisfactory ligand than [3H]mecamylamine, since it possessed a 50-fold greater affinity and its binding was far less sensitive to inorganic ions and Tris.

Expression and characterization of the rat alpha 4 beta 2 neuronal nicotinic cholinergic receptor in baculovirus-infected insect cells.

Baculovirus expression systems have been developed to generate 1) a neuronal nicotinic cholinergic receptor comprising both the alpha 4 and beta 2 subunits and 2) the alpha 4 and beta 2 subunits individually. The presence of the alpha 4 and beta 2 genes in the various baculovirus-infected Sf9 cells was confirmed following polymerase chain reaction (PCR) of the extracted viral DNAs, gel electrophoresis, and double strand sequencing. Autofluorography, following sodium dodecyl sulfate-polyacrylamide gel electrophoresis of infected cell lysates radiolabeled with 35S-methionine and immunoprecipated with mAb 270 (specific for the beta 2 subunit), revealed the presence of characteristic 52-kD bands in beta 2- and alpha 4 beta 2 recombinant viral-infected cells, but not in control cells or cells infected with wild-type virus or recombinant virus containing alpha 4 alone.

Cocaine: mechanism of inhibition of a muscle acetylcholine receptor studied by a laser-pulse photolysis technique.

Effects of cocaine on the muscle nicotinic acetylcholine receptor were investigated by using a chemical kinetic technique with a microsecond time resolution. This membrane-bound receptor regulates signal transmission between nerve and muscle cells, initiates muscle contraction, and is inhibited by cocaine, an abused drug. The inhibition mechanism is not well understood because of the lack of chemical kinetic techniques with the appropriate (microsecond) time resolution.

Effects of nicotinic dimethylaminoethyl esters on working memory performance of rats in the radial-arm maze.

Nicotine has been found to improve memory performance in a variety of tests, including the radial-arm maze. This improvement, together with the consistent finding of a decline in cortical nicotinic receptor concentration in Alzheimer's patients, has fueled the search for novel nicotinic ligands with therapeutic potential. In the current studies, a series of nicotinic compounds was tested for effects on working memory performance in the radial-arm maze.

Antagonism of nicotine's action by cocaine analogs.

Structure-activity relationships of a number of synthetic cocaine analogs are described comparing their effectiveness in antagonizing the behavioral effects of nicotine in mice with their ability to compete for [3H]mecamylamine, [3H]nicotine, and [3H]3-quinuclidinylbenzilate ([3H]QNB) binding to calf brain membranes. Within a series of phenyltropane carboxylic acid methyl esters the most potent analogues were the 4-I and 4-F-phenyl analogs, while replacement of F by Cl or alkyl groups diminished potency. The isopropyl and phenylcarboxylic acid esters were comparable in potency to the methyl esters.

Differences in ligand binding and phosphoinositide turnover between M1 muscarinic receptor gene transfected cells and mouse and rat brain membranes.

The present study describes some unexpected receptor mediated effects of N-methylcarbamylcholine on mouse M1 muscarinic receptor gene transfected cell line (M1Y1) that were not evident from biochemical studies with mouse and rat brain tissue where N-methylcarbamylcholine exhibited only nicotinic properties. Although N-methylcarbamycholine was devoid of muscarinic properties in mouse and rat brain preparations, as determined by phosphoinositide turnover and inhibition of [3H]QNB binding, it exhibited significant muscarinic characteristics in the transfected M1Y1 cell line. At a concentration of 10(-6) M or greater, N-methylcarbamycholine caused a transient increase in intracellular Ca2+ of 50 s duration that was reversible by atropine or pirezepine.

5-Isothiocyanonicotine: a high-affinity irreversible ligand for brain nicotinic receptors.

A newly synthesized affinity ligand, (R,S)-5-isothiocyanonicotine (ISCN-N) was found to inhibit irreversibly the binding of [3H]methylcarbamylcholine (a specific nicotinic receptor ligand) to brain membranes. Plots of percent inhibition versus ligand concentration yielded an IC50 of 7 x 10(-8) M for SCN-N and Ki values of 6 x 10(-9) and 2 x 10(-9) M for (R,S)-5-aminonicotine and (S)-nicotine, respectively. The IC50 value for irreversible inhibition of [3H]methylcarbamylcholine by SCN-N was 2 x 10(-7) M.

Affinity ligands and related agents for brain muscarinic and nicotinic cholinergic receptors.

This study describes the chemical synthesis and receptor binding characteristics of various affinity ligands and related ligands for brain muscarinic and nicotinic cholinergic receptors, including the 4-bromoacetamidobenzoic acid esters of dimethylaminoethanol (DMBAB) and choline (BABC) and 4-iodoacetamidobenzoylcholine (IABC). The reversible binding of [3H]3-quinuclidinylbenzilate ([3H]QNB) to calf brain membranes was inhibited in a concentration-dependent and saturable manner by DMBAB, BABC, and IABC with Ki values of 8 x 10(-7), 3 x 10(-7) and 8 x 10(-7) M, respectively; the Ki values for inhibition of reversible binding of the nicotinic ligand, [3H]methylcarbamylcholine ([3H]-MCC), were 1 x 10(-6), 6 x 10(-8), and 1 x 10(-6) M, respectively. The Ki values for irreversible inhibition of [3H]QNB binding were 8 x 10(-7), 1 x 10(-7), and 2 x 10(-7) M for DMBAB, BABC, and IABC, respectively, and for [3H]MCC binding, 8 x 10(-5), 1 x 10(-5), and 2 x 10(-5) M, respectively.

Opioid receptor antagonist affinity ligands: 6 beta-bromoacetamido-6-desoxynaltrexone and 6 beta-thioglycolamido-6-desoxynaltrexone.

The present study, utilizing thioglycolamido as the reactive group, describes the synthesis and pharmacology of a new opioid antagonist affinity ligand, 6 beta-thioglycolamido-6-desoxynaltrexone (TAN) and compares TAN with a related known compound, 6 beta-bromoacetamido-6-desoxynaltrexone (BAN). Both compounds were tested for their reversible and irreversible inhibition of [3H]naloxone binding to calf brain membranes. Reversible binding of BAN and TAN had Ki values of 1 x 10(-9) and 1 x 10(-10) M, respectively as determined by log probit plots.

4-Bromoacetamidoprocaine: an affinity ligand for brain muscarinic and nicotinic cholinergic receptors.

This study describes the synthesis, receptor binding characteristics, and some behavioral effects of p-bromoacetamidoprocaine (BAP), a new affinity ligand for brain muscarinic and nicotinic cholinergic receptors. The reversible binding of [3H]QNB to rat brain membranes was inhibited in a concentration dependent and saturable manner by both procaine and BAP, with Ki values of 4 x 10(-6) and 3 x 10(-7) M, respectively, and complete inhibition at 1 x 10(-5) M. Both procaine and BAP, although at much concentrations, inhibited the binding of [3H]methylcarbamylcholine in a concentration dependent manner, with Ki values of 5 x 10(-5) and 1 x 10(-5) M, respectively, and complete inhibition for both at 1 x 10(-3) M.

[3H]mecamylamine binding to rat brain membranes. Studies with mecamylamine and nicotine analogues.

Mecamylamine, an antagonist to nicotine, does not compete at the nicotinic recognition site, but is believed to block the ion channel of the nicotinic receptor. The present study demonstrates specific, saturable [3H]mecamylamine binding in rat brain membranes. [3H]Mecamylamine binding was destroyed by heating at 100 degrees and trypsin.

Arginine vasopressin-anti-idiotypic immunostaining of human brain cells.

Abstract Polyclonal anti-idiotypic antibodies, generated against the IgG fraction of antisera to arginine vasopressin (AVP), were shown to recognize two proteins in rat brain and bovine pituitary associated with [(3) H]AVP binding. Immunochemical analyses with these antisera revealed reactivity in paraventricular and supraoptic nucleus neuronal elements and in terminals of the posterior pituitary in the human central nervous system. With the use of a dual immunocytochemical staining technique employing both the anti-idiotype and idiotype for AVP it was possible to demonstrate a pattern of AVP-anti-idiotypic-immunoreactivity on AVP neuronal elements which suggests the existence of autoreceptors.

Differential desensitization of muscarinic receptor-stimulated phosphoinositide turnover in the rat brain.

Desensitization of the muscarinic receptor-mediated phosphoinositide (PI) turnover response in the striatum and cortex of the rat brain was examined. The rate of accumulation of inositol phosphates during carbachol-stimulated PI turnover was constant for at least 60 min in the cortex, but decreased with time in the striatum. The effects of preincubation in the presence of the muscarinic agonist carbachol on muscarinic receptor-mediated PI turnover in slices and subsequent receptor binding in cell aggregates prepared from the slices were measured.

Sites, mechanisms, and structural characteristics of the brain's nicotine receptor.

Studies are described dealing with the molecular features of nicotine, the receptor binding and psychotropic properties of nicotine agonists and antagonists, and the neuroanatomical locus of action of nicotine associated with its psychotropic action. Bridged analogues of nicotine have been developed to define the optimal conformation of the molecule for maximal receptor affinity and psychotropic action in rats. With another series of analogues, it was demonstrated that contraction of the pyrrolidine ring to a 4-member azetidine enhances potency while expansion diminishes it.

Interaction of putative vasopressin receptor proteins of rat brain and bovine pituitary gland with an antibody against a nanopeptide encoded by the reverse message of the complementary mRNA to vasopressin.

An antibody directed against the reverse message of the complementary mRNA for arginine vasopressin was demonstrated to be immunoreactive with 62 and 55 kdalton proteins, obtained by affinity chromatography of rat brain and bovine posterior pituitary extracts and believed to be associated with the vasopressin binding site. A similar pattern of immunoreactivity was seen with an anti-idiotypic antibody for arginine vasopressin.

Interaction of putative vasopressin receptors in rat brain and bovine pituitary gland with a vasopressin anti-idiotype antibody as revealed by immunoblotting.

Vasopressin (AVP)-binding proteins were obtained from rat brain and the anterior and posterior lobes of bovine pituitary glands by (a) preparation of crude membranes, (b) solubilization of membrane proteins, (c) passage through an affinity column containing immobilized AVP, and (d) elution from the column with excess AVP. Gel electrophoresis revealed protein bands of 55 and 62 kilodaltons in rat brain, bovine posterior lobe and, to a far lesser extent, in the anterior lobe, which were similar to those previously identified in rat brain to be associated with AVP binding. Immunoblotting demonstrated that the 55 kilodalton bands of rat brain and bovine pituitary gland were selectively immunoreactive with an AVP anti-idiotype antibody.

Inhibition of phosphoinositide turnover by selective muscarinic antagonists in the rat striatum. Correlation with receptor occupancy.

In the rat corpus striatum, receptor occupancy and the inhibition of phosphoinositide turnover by muscarinic antagonists have been examined under very similar conditions with respect to tissue preparation and buffer composition. The results suggest a good correlation between receptor occupancy and inhibition by muscarinic antagonists, of the carbachol-stimulated turnover of inositol phospholipids, measured by the accumulation of [3H]inositol phosphates in the presence of 5 mM LiCl. In the presence of 10 mM carbachol (CCh), the accumulation of labeled inositol phosphates was increased 8-fold above basal levels (EC50 = 95 microM).