Sympathetic nervous control of blood flow in the gills of the Atlantic cod,Gadus morhua

Summary The effects of sympathetic nerve stimulation, adrenaline and isoprenaline on the inflow pressure and efferent arterial and venous flow rates were studied in a cod gill preparation perfused at constant flow rate.The dominant effect of adrenaline was a reduced inflow pressure, accompanied by an increase in arterial flow and a decrease in venous flow. Isoprenaline also decreased the inflow pressure, but the changes in both outflow rates were small or absent.Sympathetic nerve stimulation gave arterial and venous flow changes comparable to the adrenaline effects, but the inflow pressure increased during nerve stimulation. Propranolol has little effect on the nerve responses, but phentolamine abolished or reversed the increase in inflow pressure, and also decreased or abolished the changes in outflow rates.The possible sites of action of the sympathetic fibres, and the distribution of adrenoceptors in the effector tissue is discussed. It is concluded that the main effect of sympathetic nerve stimulation is -adrenoceptor mediated, involving constriction of the arterio-venous pathway. The-adrenoceptor mediated control of total branchial vascular resistance may largely depend on circulating catecholamines.     

Sympathetic nervous control of the iris sphincter of the atlantic cod,Gadus morhua

Summary The autonomic nervous control of the cod iris has been studied. The pharmacological properties of the smooth muscles of the iris have been elucidated by agonist/antagonist studies on isolated strip preparations. Electrical stimulation of parasympathetic and sympathetic pathways to the eye have been carried out, with recordings of the movements of the iris margin. Additions of cholinergic and adrenergic antagonists in selective concentrations were made to investigate the nature of the autonomic nerve fibres controlling the iris.Isolated strip preparations of the iris sphincter contracted in response to cholinergic or-adrenoceptor agonists. There appear to be no radial muscular elements in the cod iris. The effect of carbachol on the iris sphincter could be competitively antagonized by atropine, suggesting the presence of muscarinic receptors of the smooth muscles. The effect of adrenaline was similarly antagonized by phentolamine. The effect of phentolamine, and the order of potency for the adrenergic agonists, shows the presence of-adrenoceptors in the iris sphincter.-adrenoceptors of minor importance are also suggested by the inhibitory effects of isoprenaline on preparations pre-contracted by carbachol.The indirectly acting adrenergic agonist tyramine also contracts the isolated sphincter preparations. This effect is probably due to release of nervously stored catecholamines, since tyramine lacks effect on preparations from animals pre-treated with 6-hydroxydopamine. Preparations from 6-hydroxydopamine pre-treated animals also show a 10-fold increase in the affinity for adrenaline, demonstrating the development of a pre-synaptic supersensitivity due to the destruction of adrenergic nerve terminals of the iris. Stimulation of the sympathetic chain or ciliary nerves produces a constriction of the pupil of the same side. Application of selective concentrations of the antagonists atropine and phentolamine shows that the sympathetic constrictory innervation is solely adrenergic. In some preparations a small pupillo-dilatory effect of nerve stimulation is evident after the constrictory effect has been abolished by phentolamine. This inhibitory effect can be abolished by propranolol, indicating the presence of a-adrenoceptor mediated inhibitory control of minor importance. Stimulation of the oculomotor nerve produces no consistent responses of the cod iris.Illumination of one eye produces a pupilloconstriction comparable to that seen after sympathetic nerve stimulation. The light induced response is insensitive to atropine, phentolamine and tetrodotoxin, showing a direct effect on the smooth muscles of the sphincter. There is no consensual reflex in the cod.I wish to thank Dr. Susanne Holmgren for critically examining the original draft of this paper, and Mrs. Lena Utter for skilled assistance with isolated strip preparations and processing of concentration-response data. The fish was kindly supplied by Mr. Ingmar Hakemar. This work has been supported by grants from the Swedish Natural Science Research Council, the M. Bergvall Foundation and the Adlerbert Foundation.     

Nervous control of the branchial vascular resistance of the Atlantic cod,Gadus morhua

Summary The effects on branchial vascular resistance of electrical stimulation of the nervous supply to the gills of the Atlantic cod were studied in constant pressure perfused gill preparations.Stimulation of the right sympathetic chain immediately anterior to the coeliac ganglion produces either a -adrenoceptor mediated decrease in branchial vascular resistance of the gill arches on the right side, or an -adrenoceptor mediated increase which is reversed by phentolamine to a -adrenoceptor mediated decrease in branchial vascular resistance.Stimulation of the entire vago-sympathetic nerve trunk to the third isolated gill arch produces an increase in branchial vascular resistance, which in some preparations can be reversed by atropine to a -adrenoceptor mediated decrease. A second type of constrictory innervation of vagal origin (non-adrenergic, non-cholinergic) may be concluded from the lack of blocking capacity of cholinergic and adrenergic antagonists.It is concluded that the branchial vascular bed of the cod is controlled by both sympathetic (dilatory and sometimes also constrictory) and parasympathetic (constrictory) fibres. The site of action of the nerve supply on the various effectors of the complex vasculature of the gills is not known. An autonomic innervation with its direct, rapid and restricted effects may reinforce the more general effects of circulating vaso-active substances.     

Enhancement of epidermal growth factor receptor expression on glioma cells by recombinant tumor necrosis factor α

Summary Recombinant tumor necrosis factor (rTNF; optimal dose 1000 U/ml) significantly increased the density of epidermal growth factor receptor (EGF-R) in three of four glioma cell lines in culture as determined by binding analysis of anti-EGF-R monoclonal antibody (mAb) 425. Since enhancement of EGF-R expression by rTNF- was inhibited when cells were treated with the protein synthesis inhibitor cycloheximide, the effects of rTNF may be protein-synthesis-dependent. The dose of rTNF that was optimal for up-regulation of EGF-R on glioma cells did not inhibit the growth of these cells.¹²⁵I-labeled mAb 425 lysed glioma cells in culture following its internalization into the cells. After glioma cells had been treated with rTNF, the growth-inhibitory effects of the mAb were significantly enhanced, probably a reflection of the increase in EGF-R density on the tumor cell surfaces. The rTNF effects were specific to the EGF-R and did not affect unrelated glioma-associated antigens. In our previous clinical trials,¹²⁵I-labeled mAb 425 showed immunotherapeutic effects in glioma patients. The present study provides the basis for considerations of combined immunotherapy of glioma patients with¹²⁵I-labeled mAb 425 and rTNF.     

Contribution of α-adrenoceptors to the contractility of the human myocardium in chronic coronary heart disease

The inotropic response of isolated myocardial strips to ₁-adrenoceptor stimulation was compared for patients with chronic coronary heart disease (CHD) and patients with WPW syndrome. The ₁-adrenoceptors were stimulated with 1 × 10^– M phenylephrine after blocking of the -adrenoceptors with 3 × 10^–1 M propranolol. The inotropic activity was recorded in the isometric mode. In the myocardium without signs of ischemic damage, stimulation of the ₁-receptors caused a slowly developing single-phase positive inotropic response. The myocardium of the CHD patients was characterized by a three-phase response. The specific features of the inotropic response to ₁-adrenoceptor stimulation in the CHD patients were assumed to be determined by changes in intracellular homeostasis of Ca²⁺. Electromechanical coupling in cardiac myocytes of CHD patients depends on Ca²⁺ deposited in the sarcoplasmic reticulum to a greater extent than coupling in the intact myocardium. An additional positive inotropic effect is possible upon exogenous calcium influx into cardiac myocytes.Translated from Fiziologiya Cheloveka, Vol. 31, No. 1, 2005, pp. 133–136.Original Russian Text Copyright © 2005 by Afanasev, Ugdyzhekova, Karpov.     

Release of [3H]noradrenaline by α1-adrenoceptor agonists

Mouse isolated vas deferens preincubated with [³-H]noradrenaline was superfused and the effect of ₁-adrenoceptor agonists was studied on the release of total radioactivity ([³H]noradrenaline +³H-metabolites) and [³H]noradrenaline. Reverse phase high pressure liquid chromatography (HPLC) combined with scintillation spectrometry was used to separate [³H]noradrenaline from its metabolites. Among the ₁-adrenoceptor agonists (1-phenylephrine, ST-587(2-(2-chloro-5-trifluoromethyl phenylimino)-imidazole), (–)-amidephrine, methoxamine, cirazoline and l-noradrenaline) studied l-phenylephrine, ST-587 and l-noradrenaline were capable of releasing³H-noradrenaline. The effect of noradrenaline was stereospecific. As determined by HPLC combined with scintillation spectrometry the release of total radioactivity in response to l-noradrenaline is mainly due to [³H]noradrenaline. It is suggested that l-noradrenaline, l-phenylephrine, and ST-587 in addition to their direct effect on different receptors they also have indirect action through the release of noradrenaline which might be partly involved in the pharmacological responses. The mechanisms whereby l-noradrenaline and l-phenylephrine release noradrenaline would appear to involve a saturable Ca-independent and a cocaine and temperature sensitive process. On the basis of our findings among the ₁-adrenoceptor agonist studied (–)-amidephrine, methoxamine and cirazoline is a better choice than l-phenylephrine or ST-587 for selective stimulation of postjunctional ₁-adrenoceptor, they do not release noradrenaline.     

The regions of alpha-neurotoxin binding on the extracellular part of the alpha-subunit of human acetylcholine receptor

A set of 18 synthetic uniform overlapping peptides spanning the entire extracellular part (residues 1–210) of the -subunit of human acetylcholine receptor were studied for their binding activity of¹²⁵I-labeled -bungarotoxin and cobratoxin. A major toxin-binding region was found to reside within peptide 122–138. In addition, low-binding activities were obtained with peptides 34–49 and 194–210. It is concluded that the region within residues 122–138 constitutes a universal major toxin-binding region for acetylcholine receptor of various species.     

TNF/LTα double knockout mice display abnormal inflammatory and regenerative responses to acute and chronic liver injury

Following acute liver injury, hepatocytes divide to facilitate regeneration. However, during chronic injury, hepatocyte proliferation is typically blocked and repair is mediated through liver progenitor (oval) cells. Signalling of the p55 tumour necrosis factor (TNF) receptor is central to these processes. Two ligands for p55 are known: TNF and lymphotoxin-alpha (LT). However, one study suggests that another exists that mediates liver injury following viral challenge. We have therefore investigated whether ligands other than TNF and LT are required for liver regeneration following either acute or chronic injury. Wild-type and double TNF/LT knockout (TNF^–/–LT^–/–) mice were subjected to either partial hepatectomy (PHx) or a choline-deficient ethionine-supplemented (CDE) diet. Proliferating hepatocytes, oval cells and inflammatory cells were identified and quantified in liver sections by immunohistochemistry. Liver inflammatory cells were characterised by cell surface antigen expression. Liver damage and mortality were monitored. Both hepatocyte and oval cell proliferation was reduced in TNF^–/–LT^–/– mice. Lymphocyte clusters were evident in all TNF^–/–LT^–/– livers and were heterogeneous, comprising B and T lymphocytes. PHx evoked liver inflammation in TNF^–/–LT^–/– but not wild-type mice, whereas no difference was apparent between genotypes in CDE experiments. Thus, TNF/LT signalling mediates liver regeneration involving both hepatocytes and progenitor cells. The hyper-inflammatory response following PHx in TNF^–/–LT^–/– animals, which is absent following CDE-induced injury, demonstrates that the two forms of liver injury evoke discrete inflammatory responses and provides a model in which such differences can be examined further.     

Alpha-1 and alpha-2 adrenoceptor binding in cerebral cortex: Role of disulfide and sulfhydryl groups

The triated adrenergic antagonists Prazosin ([³H]PRZ) and Idazoxan ([³H]IDA, or RX-781094) bind specifically and with high affinity to ₁ and ₂-adrenoceptors respectively, in membrane preparations from cerebral cortex. Saturation experiments performed to determine the density of receptors and the dissociation constant (K _d) were analyzed by the methods of Eadie Hofstee, iterative modelling, and the procedure of Hill, while the specificity of the labelling was verified by displacement experiments. Since receptors are proteins, we examined the role of disulfide (–SS–) bridges and sulfhydryl (–SH) groups in the specific combination of [³H]PRZ and [³H]IDA to the ₁ and ₂-adrenoceptors. Pretreatment of the membranes with the –SS– reactive DL-dithiothreitol (DTT) or the alkylating agent N-ethylmaleimide (NEM), alone or in combination, decreased specific binding of both ligands, with only minor changes in the non-specific counts. The [³H]IDA binding (₂-sites) was more sensitive to both DTT and NEM than the [³H]PRZ sites (₂-adrenoceptors), and the initial changes induced by alkylation of the ₂-site were due to an important decrease in the affinity for [³H]IDA, as judged by the increase in theK _d. This modulation in the affinity caused by alkylation of a thiol group could explain the higher potency of the blocking agent tetramine disulfide benextramine at the ₂-site. The results provide evidence for the participation of –SS– and –SH groups in the binding site of ₁ and ₂-adrenoceptors in the cerebral cortex.     

The use of 1JCαHα coupling constants as a probe for protein backbone conformation

Summary Simple pseudo-3D modifications to the constant-time HSQC and HCACO experiments are described that allow accurate (±0.5 Hz) measurement of one bond J_CH coupling constants in proteins that are uniformly enriched with ¹³C. An empirical ,-surface is calculated which describes the deviation of ¹J_CH from its random coil value, using 203 ¹J_CH values measured for residues in the proteins calmodulin, staphylococcal nuclease, and basic pancreatic trypsin inhibitor, for which and are know with good precision from previous X-ray crystallographic studies. Residues in -helical conformation exhibit positive deviations of 4–5 Hz, whereas deviations in -sheet are small and, on average, slightly negative. Data indicate that ¹J_CH depends primarily on , and that ¹J_CH may be useful as a qualitative probe for secondary structure. Comparison of ¹J_CH coupling constants measured in free calmodulin and in its complex with a 26-aminoacid peptide fragment of myosin light-chain kinase confirm that the calmodulin secondary structure is retained upon complexation but that disruption of the middle part of the central helix is even more extensive than in free calmodulin. Supplementary material available from the authors: One table listing 352 ¹J_CH and ¹J-values, together with ,-values for 203 residues of known conformation. Two figures showing (a) a Ramachandran plot of the ,-values of 203 residues used in deriving ¹J(,), and (b) the r.m.s.d. ¹J(,) distribution.     

Surfactant effects on adrenergic responses in the gills of the flounder (Platichthys flesus L.)

Summary The interactions between catecholamines and surfactants was investigated in perfused gills of the marine teleostPlatichthys flesus L. The activity of the branchial ion pumps was monitored via the electrogenic transepithelial potential (inside positive) measured in gills bathed and perfused with identical saline. Vascular resistance of the arterio-arterial and arterio-venous pathway was also recorded simultaneously by measuring respectively the afferent perfusion pressure and venous flow in gills perfused at constant flow and at constant efferent pressure. The specific effects of respective - and -adrenergic receptor stimulation was investigated by the administration of discrete doses of either adrenaline in the presence of 10 mol l^–1 propranolol or isoprenaline in the perfusate. In the absence of surfactants the -adrenergic effects were an inhibition of electrogenic ion transport, a decrease in venous flow and an increase in the vascular resistance of the arterioarterial vascular pathway. In contrast the -adrenergic effects consisted of a stimulation of electrogenic ion transport and a vasodilation of the arterio-arterial pathway. Both anionic (linear alkyl sulphonate; sodium lauryl sulphate) and non-ionic (nonyl phenol ethoxylate; synthetic alcohol ethoxylate) surfactants were administered in the perfusate at nominal concentrations of 1 mol l^–1 (0.3–0.5 mg l^–1). None of these compounds had any effect on the affinity or the efficacy of the -adrenergic responses. In contrast there was a significant reduction in the efficacy of isoprenaline in the presence of all of the surfactants used but only in the case of the synthetic alcohol ethoxylate was there an effect on the affinity of this agonist for the -adrenergic receptor. The results are discussed in the context of the mechanism of action of these environmental contaminants and the nature of adrenergic receptors in the gill.     

Isolation and characterization of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by the mating hormone a-factor

Summary Nine independent mutants which are supersensitive (ssl ^–) to G1 arrest by the mating hormone a-factor were isolated by screening mutagenized Saccharomyces cerevisiae MAT cells on solid medium for increased growth inhibition with a-factor. These mutants carried lesions in two complementation groups, ssl1 and ssl2. Mutations at the ssl1 locus were mating type specific: MAT ssl1 ^– cells were supersensitive to -factor but MAT ssl1 ^– were not supersensitive to -factor. In contrast, mutations at the ssl2. locus conferred supersensitivity to the mating hormone of the opposite mating type on both MAT, and MATa cells. The -cell specific capacity to inactivate externally added a-factor was shown to be lacking in MAT ssl1 ^– mutants whereas MAT ssl2. cells were able to inactivate a-factor. Complementation analysis showed that ssl2 and sst2, a mutation originally isolated as conferring supersensitivity to -factor to MATa cells, are lesions in the same gene. The ssl1 gene was mapped 30.5 centi-Morgans distal to ilv5 on chromosome XII.     

Analysis of exposed regions on the main extracellular domain of mouse acetylcholine receptor α subunit in live muscle cells by binding profiles of antipeptide antibodies

To study the structural organization of the main extracellular domain of the nicotinic acetylcholine receptor (AChR) subunit in live muscle cells, we examined the native membrane-bound receptors in cultured mouse skeletal muscle cells for their ability to bind a panel of antibodies against uniform-sized overlapping synthetic peptides which collectively represent this entire domain. The binding profile indicated that the regions 23–49,78–126,146–174, and182–210 are accessible to binding with antibody. Residues23–49,78–126, and194–210 contain binding regions for-neurotoxin and some myasthenia gravis autoantibodies. A comparison of this binding profile with the profile obtained for membrane-boundTorpedo californica AChR in isolated membrane fractions showed some similarities as well as significant differences between the subunit organization in the isolated membrane fraction and that in the membrane of live muscle cells. Regions89–104 and158–174, which are exposed in the isolated membrane fraction, are also exposed in the live cell. On the other hand, regions23–49, and182–210, which are exposed in the live cell, are not accessible in the isolated membrane and, furthermore, the region1–16, which has marginal accessibility in the cell, becomes highly accessible in the membrane isolates. The exposed regions defined by this study may be the primary targets for the initial autoimmune attack on the receptors in experimental autoimmune myasthenia gravis.     

Effects of catecholamines on isolated systemic and branchial vascular beds of the cod,Gadus morhua

Summary The effects of catecholamines on the vascular resistance of the gills, the gas gland and the tail of the Atlantic cod, were studied in isolated preparations perfused at a constant pressure. Adrenaline caused a biphasic response of the gill vasculature with an initial constriction followed by a dilation. The noradrenaline response was usually dilatory, but constrictory responses at all or some concentrations were occasionally seen in some specimens. The branchial dilation was caused by adrenaline concentrations even lower than those found in normal cod plasma (30 nM). The gas gland and the tail vasculature were constricted by both adrenaline and noradrenaline. A flow reduction in the isolated preparations of about 10% at concentrations of adrenaline equalling that in normal cod plasma (30 nM) was increased to about 30% (tail) or 45% (gas gland) at concentrations of adrenaline similar to that found in cod plasma during stress (300 nM).The constrictory responses to adrenaline and noradrenaline were antagonized in all preparations by the alpha adrenoceptor antagonist, phentolamine (10^–6 M) and the dilatory response of the branchial vasculature to these agonists was reduced or abolished by propranolol (10^–6 M).It is concluded that adrenaline, in the concentrations found in cod plasma at rest and after stress, has pronounced effects on the organs studied, especially the gill vasculature, and may thus contribute to the overall control of the circulatory system. The branchial vasculature will also be affected by changes in noradrenaline concentration which occur in animals at rest and under stress.     

Ontogeny of estrogen receptor (ER) alpha and its co-localization with pituitary hormones in the pituitary gland of chick embryos

Estrogen is involved in regulating the development and hormone secretion of the anterior pituitary gland following its binding to estrogen receptors (ERs) expressed on pituitary cells. However, the pituitary is comprised of several cell types, and to date, there is no data about the specific cell types expressing ERs in embyonic chick pituitary. We therefore followed, by immunohistochemistry, the ontogeny of the pituitary ER alpha (ER), and the cell types expressing ER throughout chick embryo development. ER immunoreacitivity was restricted to the nuclei of pituitary cells. ER-immunopositive (ER⁺) cells were first detected at embryonic day 6.5 (E6.5), after which ER⁺ cells were consistently detected throughout the anterior pituitary gland, although the density of ER⁺ cells in the caudal lobe of the pars distalis was higher than that in the cephalic lobe. The proportion of ER⁺ cells in the pituitary was about 6% at E8.5; expression increased to 22% by E18.5 of gestation, with no additional change until hatching. Double-labeling of ER and pituitary hormones showed that the dominant cell types expressing ER were gonadotrophs immunopositive for luteinizing hormone (LH); the proportion of ER⁺ cells expressing LH increased throughout gestation and reached approximately 57% at hatching. About 2%–6% of thyroid-stimulating-hormone-immunopositive and 1%–2% prolactin-immunopositive cells expressed ER at later stages of embryonic development, but no growth-hormone-positive or adrenocorticotropic-hormone-positive cells expressed ER during the embryonic period. Thus, gonadotrophs are the main cell population expressing ER in the anterior pituitary gland of chick embryo, and ER is involved in regulating the development of the pituitary gland and the maturation of the hormone-secreting function.This work was supported by grants from the Natural Science Foundation for Outstanding Young Scientists of China (30325034) and the Natural Science Foundation of China (30170693, 30471264).     

Enhancement of production of cloned α-amylase by lactic acid feeding from recombinant Saccharomyces cerevisiae using a SUC2 promoter

-Amylase production was higher (13 units ml^–1) when a recombinant Saccharomyces cerevisiae containing a SUC2 promoter was grown with 10 g lactic acid l^–1 than without addition (8 units ml^–1). With continuous lactic acid feeding in the inducing phase, -amylase increased to 79 units ml^–1 in a 1-l jar fermenter.     

Adrenoceptor heterogeneity in the ruminal epithelium of sheep

The pre-gastric rumen of sheep plays a crucial role in the fermentation of nutrients and in the absorption of nutrients and minerals. Adrenaline has been shown previously to increase ruminal absorption of glucose and water. The present study was intended to elucidate whether ruminal ion transport is also altered by adrenaline. In Ussing chambers, changes of I_sc were recorded in isolated ovine ruminal epithelia after the serosal additions of adrenoceptor agonists or antagonists. I_sc increased after the addition of adrenaline (10^–4 M) or clonidine (₂-agonist, 10^–4 M), but decreased after the addition of isoproterenol (-agonist, 10^–4 M) or terbutaline (₂-agonist, 10^–5 M). The effect of adrenaline on I_sc was augmented by the adrenoceptor antagonists prazosin (₁, 10^–4 M) and bupranolol (, 10^–6 M), but inversed by yohimbine (₂, 10^–5 M). Adrenaline induced an increase in Na⁺ net flux across the epithelium that was larger than the increase in equivalent current flow. It is concluded that adrenaline differentially regulates ion transport across the ruminal epithelium via ₁-, ₂-, and ₂-receptors. The main effect is a stimulation of electroneutral and electrogenic Na⁺ absorption. This stimulated Na⁺ absorption might be causative of increased water absorption from the rumen as described previously.     

Manifestation of intramolecular motions on pico- and nanosecond time scales in 1H−15N NMR relaxation: Analysis of dynamic models of one- and two-helical subunits of bacterioopsin

Summary The influence of the internal dynamics of two polypeptides comprising transmembrane -helix A or two -helices A and B of bacterioopsin on experimentally accessible ¹⁵N NMR relaxation rates was investigated by molecular dynamics (MD) simulations, combined with more simple mechanic considerations. Model-free order parameters and correlation times of internal motions [Lipari, G. and Szabo, A. (1982) J. Am. Chem. Soc., 104, 4546–4559] were calculated for these models. It was found that both peptides exhibit two types of internal motions of the amide bonds, on the pico- and nanosecond time scales, affecting ¹⁵N NMR relaxation. The fast fluctuations are local and correspond to the librational motions of the individual N–H vectors in an effective potential of atoms of the surrounding matrix. In contrast, the motions on the nanosecond time scale imply concerted collective vibrations of a large number of atoms and could be represented as bending oscillation of -helices, strongly overdamped by the ambient solvent. A few other molecular mechanisms of slow internal motion were found, such as local distortions of the -helices (e.g., -aneurysm), delocalized distortions of the -helical backbone, as well as oscillations of the tilt angle between the axes of the -helices A and B. The results are compared with ¹⁵N NMR relaxation data measured for the (1–36)bacterioopsin and (1–71)bacterioopsin polypeptides in chloroform-methanol (1:1) and in SDS micelles [Orekhov, V.Yu., Pervushin, K.V. and Arseniev, A.S. (1994) Eur. J. Biochem., 219, 887–896].Abbreviations C2 baeterioopsin-(7–63)-peptide - sA bacterioopsin-(7–32)-peptide - CPMG Carr-Purcell-Meiboom-Gill - MD molecular dynamics - rmsd root-mean-square deviation     

Regulation of phagocytic process of macrophages by noradrenaline and its end metabolite 4-hydroxy-3-metoxyphenyl-glycol. Role of α- and β- adrenoreceptors

The regulatory capacity of noradrenaline and its end metabolite 4-hydroxy-3-metoxyphenylglycol (HMPG) on the complete phagocytic process of macrophages were investigated. Either noradrenaline or HMPG did not modify adherence. However, 10^–12 M of noradrenaline stimulated the chemotaxis of macrophages, mainly mediated by -adrenergic receptors. In contrast, 10^–12 M of HMPG induced an opposed effect on this stage of the phagocytic process. To stimulate phagocytosis, it is necessary to employ a higher concentration (10^–5 M) of noradrenaline and this effect was blocked with either 10^–6 M propranolol or 10^–6 M phentolamine, and maintained by HMPG. Noradrenaline and HMPG did not modify the microbicide capacity of macrophages (measured by O₂ ^– production after phagocytosis). In conclusion, noradrenaline modulates the phagocytic process of macrophages, and this modulation is completed by HMPG, maintaining the phagocytic functions at physiologically optimal levels. Modulation of chemotaxis is mainly mediated by a-receptors and phagocytosis needs both - and -receptor-stimulation.     

A new 3D HN(CA)HA experiment for obtaining fingerprint HN-Hα cross peaks in15N- and13C-labeled proteins

Summary A new 3D¹H–¹⁵N–¹³C triple resonance experiment is presented that provides in-phase absorptive cross peaks between amide protons and -protons of the same and the sequentially preceding residue. The experiment yields similar connectivities as those described previously by Montelione and Wagner (1990a) [J. Magn. Reson.,87, 183–188] and Kay et al. (1991) [J. Magn. Reson.,91, 84–92]. However, the pulse sequence was designed to minimize the time that transverse coherence of the¹³C nucleus is present, since this nucleus has the shortest transverse relaxation time of all the nuclei involved in these experiments. This is achieved by using a coherence transfer pathway from¹H^N to¹⁵N,¹³C,¹H and back to the¹H^N. In the sequence described, transverse¹³C coherence is present only for a length of ca. I¹J(C-H). This reduces loss of signal due to transverse relaxation. We tested the technique on uniformly¹⁵N- and¹³C-enriched T4 lysozyme.