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M Nilsson

Publications and source records attributed to M Nilsson.

At least 19 recordsLinked to original sources

[Astrocytes are of vital significance for the central nervous system].

The tissue of the CNS (central nervous system) is composed of neurons and neuroglia. Whereas neurons develop an ability for the rapid transduction of specific signals, astrocytes develop an ability to modulate the extracellular neuronal environment, and in mature CNS tissue manifest a capacity for active uptake of amino acids and ions. Astrocytes can control extracellular volume by regulation of their own volume, and are intimately involved in the neuronal exchange of trophic substances and metabolites. Astrocytic processes extend to blood vessel walls, the brain surface, the ventricular wall, neuronal cell bodies and synapses. Astrocytes are abundantly supplied with membrane receptors for various neurotransmitters, coupled to such second messenger systems as cyclic AMP (adenosine monophosphate) or the phosphatidylinositol cycle. Activation of the receptors results in changes in oxidative metabolism, cell morphology, cell volume, and immunocompetence: and recent findings have shown the occurrence of receptor-mediated changes in amino acid uptake. Thus, by modulating the extracellular environment, astrocytes can simultaneously modulate the sensitivity and/or excitability of large numbers of neurons. In the article are presented recent research findings suggesting astroglial cells to be targets for neurotransmitters, and probably to be actively involved in higher cognitive functions. Advances in our knowledge of astroglial cell characteristics might improve our understanding of behavioural disturbances and diseases of the CNS.

Astrocytes

Volume regulation of single astroglial cells in primary culture.

Relative volume variations in cultured astrocytes were examined by microspectrofluorimetry after loading the cells with the highly fluorescent intracellular probes 2,7-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF/AM) or fura-2/AM. At their isosbestic points, 450 nm and 358 nm, respectively, the probes were ion-insensitive and the fluorescent signals emitted related only to the intracellular dye concentration. By varying the excitation wavelengths, changes in intracellular pH or Ca2+ transients could be recorded simultaneously with the relative volume variations of the individual cells. After exposure to a hypotonic buffer, type 1 astrocytes swelled within 30 s and subsequently underwent regulatory volume decrease (RVD). When exposed to a hypertonic buffer, the astrocytes shrunk and exhibited regulatory volume increase (RVI). One mM glutamate induced an increase in astrocyte volume in 60 sec and evoked cytosolic Ca2+ transients but did not change intracellular pH.

Animals

Process monitoring by flow-injection immunoassay. Evaluation of a sequential competitive binding assay.

A new variation on the theme of flow-injection binding assays is presented, namely the sequential competitive binding assay, in which the sample containing the native antigen is first introduced into the flow system followed by a pulse of labelled antigen. The flow-injection binding assay was used in monitoring the effluent from column chromatographic separations of proteins. By applying a computer-based evaluation system, concentrations are automatically read and compensation is made for denaturation in the affinity sorbent.

Binding, Competitive

Receptor-coupled uptake of valproate in rat astroglial primary cultures.

Various receptor ligands were investigated for their effects on the uptake of the antiepileptic drug valproic acid (VPA) in primary astroglial cultures from cerebral cortex of neonatal rats. After stimulation with the alpha 1-adrenoceptor agonist phenylephrine, 5-hydroxytryptamine (5-HT) or the glutamate receptor agonists glutamate, quisqualate and kainate, the Vmax and Km values for the drug transport increased. On the contrary, after exposure to the alpha 2-adrenoceptor agonist clonidine, Vmax and Km decreased. The effects were reversed in comparison to the control level in the presence of selective receptor antagonists. The data indicate a specific coupling between receptors and the uptake system for VPA. Furthermore, the results may have significant implications, as they suggest that receptors on astrocytes can be involved in the local regulation of drug transport in brain.

6-Cyano-7-nitroquinoxaline-2,3-dione

Chronic elevation of cAMP levels induces changes in the adenylate cyclase system, opiate receptor sensitivity and levels of Gs-mRNA in cultured neurons.

Neurons in primary culture were treated (5 days) with the adenylate cyclase stimulator 10(-5) M forskolin. The basal adenylate cyclase was decreased by 57%. The acute stimulatory effect of forskolin was down-regulated by 48%. The inhibitory effects of the 3 opiate receptor agonists (mu, delta and kappa) were partly abolished. The abundance of mRNA encoding the stimulatory G-protein (Gs) was decreased prominently. The data indicate that the cAMP system in neuronal cells in primary culture is under dynamic regulation, possibly including altered Gs-protein gene expression. Furthermore, long-term forskolin treatment might induce increased proliferation in susceptible neural blast cells.

Adenylyl Cyclases

Agonist-evoked Ca2+ transients in primary astroglial cultures--modulatory effects of valproic acid.

Cytosolic Ca2+ ([Ca2+]i) activity was measured in individual type 1 astroglial cells in primary culture after exposure to glutamate (Glu), quisqualate (QA), gamma-aminobutyric acid (GABA), 5-hydroxytryptamine (5HT), and noradrenaline (NA) by using the Ca2+ indicator dye fura-2/AM in a computerized microspectrofluorimetric system. Various patterns of Ca2+ transients were observed, but the most common was biphasic, having an initial sharp peak, rising immediately after stimulation, and then declining to a lower but sustained Ca2+ level. The only substance that diverged from this pattern was GABA, which induced a Ca2+ response with longer latency and a single-phase curve. The effects of the anticonvulsive drug Na(+)-valproate (VPA) were also investigated. After both acute and chronic (5-7 days) exposure to 10(-4) M VPA, the GABA-evoked rises in [Ca2+]i were completely inhibited. VPA also had acute effects on the 5HT- and Glu-evoked Ca2+ spikes. The Ca2+ responses after 5HT stimulation were greatly reduced after exposure to 10(-4) M VPA. The responses after glutamate stimulation were, on the contrary, increased after a similar exposure. No VPA effects were seen on the curve patterns of QA and NA stimulations. The most frequent agonist-evoked responses were seen after stimulation with 5HT and NA, where over 80% of the tested cells responded. For QA and Glu, the response frequencies were about 40% each, while for GABA it was 20%. The responses after 5HT and NA stimulation were blocked to baseline levels after exposure to ketanserin (5HT2 receptor antagonist) and a combination of prazosin, yohimbine, and propranolol (alpha 1, alpha 2, and beta adrenoceptor antagonists, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Interactions between valproate, glutamate, aspartate, and GABA with respect to uptake in astroglial primary cultures.

Astrocytes have been proposed to regulate the extracellular space in the brain, even if rather little is known about their specific functions. One possibility for obtaining more knowledge on the functions of astroglial cells is to examine how they respond on exposure to pharmacological agents. Na(+)-valproate is an anticonvulsive drug which is used in the treatment of several types of epilepsy. The mechanisms of action of the drug are not fully understood, but the GABA-ergic system, both in neurons and astrocytes, has been shown to be affected. In the present study, the effects of valproate were investigated on astroglial cells in primary cultures from newborn rat cerebral cortex. The transport of the drug itself and its effects on the transport of the amino acid transmitters glutamate, aspartate and gamma-aminobutyric acid (GABA) into astrocytes were examined. The [3H]valproate transport into the astrocytes was increased after exposure to L-glutamate but not L-aspartate. On the other hand, after acute exposure for the drug, the transport of [3H]L-glutamate and [3H]L-aspartate decreased, as also did the affinity but not the transport capacity for the [3H]GABA uptake. However, after 5 days chronic valproate exposure, no effects could be seen on the uptake kinetics of L-glutamate or L-aspartate. For GABA, the affinity decreased, while the transport capacity remained unchanged compared with controls. The results showed that valproate, glutamate, aspartate and GABA were capable of interacting significantly with each others transport into the astrocytes.

Animals

The use of hyaluronidase and glucosidase to remove mucus from the rat middle ear cavities for SEM studies.

To conduct SEM studies on epithelium containing mucus-producing cells it is essential to remove the mucus which normally obscures the epithelial surface. This study presents a method which effectively removes the covering layer of mucus in the rat middle ear. Healthy Sprague-Dawley rats were decapitated and the middle ears dissected free. Incubation and agitation of the middle ear specimens in hyaluronidase (50 IE ml-1) and/or glucosidase (8%) removed the mucus from the middle ear cavity without altering the surface structures. It was also revealed that substances such as polyvinyl-pyrrolidone (PVP) (used to increase the colloid osmotic pressure of, e.g., the fixative solution) must be omitted when preparing ciliated specimens for SEM.

Animals

Structural and functional aspects of the thyroid follicular epithelium.

The thyroid epithelium is morphologically and functionally polarized, with an apical surface facing the follicular lumen containing colloid and a basolateral surface facing the interstitium. Iodination and thyroid hormone synthesis occur in the colloid at the apical plasma membrane. The introduction by Mauchamp et al. of primary cultures of porcine thyroid cells grown as a polarized, confluent monolayer on a filter in a bicameral chamber system has now made it possible to study in more detail the barrier function and vectorial ion transport in the thyroid epithelium. The follicular cells form a very tight monolayer (transepithelial resistance > 6000 ohm cm2) and establish a transepithelial potential difference (apical medium negative) of about 20 mV. These parameters are rapidly influenced by TSH, mainly by an action on apical sodium channels, and by EGF. The integrity of the barrier is, as in other epithelia, dependent on extracellular calcium. A calcium-dependent cell adhesion molecule, uvomorulin, is expressed at the lateral plasma membrane surface. EGF induces cell proliferation as well as migration of some of the epithelial cells to a position below the monolayer, which however maintains its polarity and barrier function. In contrast, during TPA-induced proliferation the barrier function is disrupted. Iodide is vectorially transported in basoapical direction while the epithelial layer is virtually impermeable for iodide transfer in the opposite direction. Iodide is concentrated in the cell by the basolateral "iodide-pump" and its efflux across the apical plasma membrane is rapidly and selectively increased by TSH via cAMP. EGF inhibits vectorial basoapical iodide transport mainly by reducing the iodide permeability of the apical plasma membrane. Together, these recent observations indicate that the ion content of the follicular lumen is strictly controlled by the thyroid epithelium.

Animals

Comparative multiple-dose pharmacokinetics of controlled-release levodopa products.

The multiple-dose (200 mg levodopa t.i.d.) pharmacokinetic profile of two controlled-release products of levodopa (Madopar HBS and Sinemet CR) was compared to conventional Madopar capsules in 18 healthy volunteers in a cross-over, randomized design. A pronounced controlled-release profile of the Madopar HBS and Sinemet CR product was demonstrated compared to conventional Madopar capsules with a significant (p < 0.001) decrease (-40 and -55%) in Cmax and a significant (p < 0.001) increase (+237 and +256%) in morning Cmin for the 200 mg t.i.d. dosage schedule. Almost equivalent bioavailability (85-90%) of levodopa was demonstrated for the controlled-release formulations relative to that of conventional Madopar capsules. The Madopar HBS formulation was bioequivalent with Sinemet CR with respect to levodopa, but it exhibited a moderately higher fluctuation index compared to Sinemet CR as a result of somewhat higher Cmax and lower Cmin values for the Madopar HBS formulation. 3-OMD (a metabolite of levodopa) levels were significantly (p < 0.05) higher for Madopar HBS and Madopar compared to Sinemet CR. The higher 3-OMD levels for the levodopa/benserazide combinations are consistent with a more potent decarboxylase inhibitory activity of benserazide as compared to carbidopa. The number of adverse events was highest for conventional Madopar (n = 18) compared to the controlled-release formulations (n = 12 for Sinemet CR and only 2 for Madopar HBS). A more efficient inhibition of dopamine formation from levodopa (resulting in higher 3-OMD levels) by Madopar HBS was consistent with the superior tolerability (especially for initial nausea) observed for the Madopar HBS formulation as compared to Sinemet CR.

Adult

Polarized efflux of iodide in porcine thyrocytes occurs via a cAMP-regulated iodide channel in the apical plasma membrane.

The intracellular regulation of thyrotropin-stimulated iodide efflux was studied in polarized porcine thyrocytes grown as a continuous, tight monolayer in bicameral culture chambers. From a previous study using this system we know that thyrotropin rapidly increases iodide efflux in the apical but not basal direction of the polarized epithelium. [125I]-iodide efflux in apical direction was stimulated by thyrotropin in a concentration-dependent manner (1-10 U/l), whereas efflux in basal direction was unchanged at any thyrotropin dose. Thyrotropin-induced elevation of intracellular cAMP showed a corresponding concentration dependence. The selective stimulation of apical efflux by thyrotropin was evident also when re-uptake of iodide released in basal direction was blocked by perchlorate. The effect of thyrotropin on apical efflux was mimicked by 8-bromo-cAMP and forskolin, whereas agents known to activate the Ca2+/phosphatidylinositol cascade (epidermal growth factor) and protein kinase C (phorbol ester) or increase cytosolic [Ca2+] (A23187) were inactive. We conclude that the selective stimulation by thyrotropin of apical iodide efflux, corresponding to efflux in luminal direction in intact follicles, occurs via cAMP-regulated iodide channels present in the apical domain of the plasma membrane.

8-Bromo Cyclic Adenosine Monophosphate

Subtraction radiography of interradicular bone lesions.

Subtraction and conventional radiography were evaluated for their diagnostic potential to assess interradicular bone lesions in the mandibular premolar region. Both conventional radiographs and subtraction images were interpreted by 10 observers. The receiver-operating characteristic (ROC) technique was used to compare the two techniques. The diagnostic validity was higher for the subtraction technique, both for lesions confined to cancellous bone and for lesions including the cortical bone, than for the conventional technique. For bone defects confined to cancellous bone the diagnostic accuracy was lower than those reported from periapical bone lesions irrespective of whether subtraction or conventional radiography was used. We conclude that subtraction radiography improves the detectability of bone lesions, shallow ones in particular. Lesions in the interradicular bone are more difficult to detect than those in the periapical bone.

Alveolar Process

The microwave oven in temporal bone research.

The microwave oven has recently been introduced in morphological research. It can be used for virtually all procedures in histological processing; fixation, dehydration, embedding, staining, immunotechniques, and the techniques used for electron microscopy. Here we report that the microwave oven histoprocedures also have several advantages in temporal bone research on animals. The microwave oven has been tested for (i) fixation, (ii) decalcification, (ii) staining of semithin plastic sections. The microwave oven procedures are time-saving and results in an excellent preservation of the tissues. The microwave histoprocessing should also be applicable to temporal bone research in human.

Animals

Adrenergic and 5-HT2 receptors on the same astroglial cell. A microspectrofluorimetric study on cytosolic Ca2+ responses in single cells in primary culture.

Noradrenaline (NA) or 5-hydroxytryptamine (5-HT) evoked cytosolic Ca2+ mobilization in single type 1 astrocytes in primary culture from the cerebral cortex of newborn rat. The Ca2+ indicator dye fura-2/AM was used in a microspectrofluorimetric system to visualize fluctuations in the intracellular Ca2+ concentration. Activation of the adrenergic receptors alpha 1, alpha 2 and beta, or activation of the 5-HT2 receptors elicited different responses of Ca2+ mobilization with different types of Ca2+ spikes or oscillations. Principally, 4 different types of Ca2+ responses could be obtained: a sharp spike, which declined back to baseline; an initial sharp spike, which declined to a smaller but sustained Ca2+ elevation; an initial sharp spike which declined and showed low amplitude oscillations; and a sharp spike which declined back to baseline with baseline oscillations. Applications of the alpha 2 adrenoceptor agonist clonidine to individual astroglial cells evoked Ca2+ transients mostly in young cultures (cultivated for 7-10 days), while applications of the alpha 1 adrenoceptor agonist phenylephrine evoked Ca2+ transients mostly in older cultures (17-21 days of cultivation). Applications of the beta adrenoceptor agonist isoproterenol evoked Ca2+ transients in both young and older cultures, however, more frequent in older cultures. The alpha 2 and beta receptor responses were dependent on external Ca2+ levels. The NA-evoked Ca2+ responses were seen in cultivated cells at all ages, but were more frequent in older cultures. Approximately 50% of the astroglial cells in 8 day old cultures responded to 5-HT with a cytosolic Ca2+ mobilization and 80% of the cells in 21 day old cultures responded.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

[Effect of alfa-2b interferon on prognostic parameters and clinical events in HIV positive patients in the LAS/ARC stage].

A number of in vivo and in vitro results suggest that interferons have an antiretroviral effect on HIV. To check this, 15 HIV-positive patients who had no full-blown AIDS, were treated with recombinant interferon alpha 2b (5 mill. IU s. c. three times a week) over a period of six months. Twelve to 16 weeks after the initiation of treatment, an increase in CD 4 lymphocytes (+16%), NK cells (+16%), lymphocytes stimulation by con A (+ 176%), neopterin (+66%), and beta-2-microglobulin (+19%) was observed. By the end of the study, all these parameters had slightly decreased again. In all patients with CD4 lymphocytes greater than 0.2 c/nl, we observed a decrease in p24 antigen levels, but in patients with CD4 lymphocytes less than 0.2 c/nl, an increase. It would thus seem that any antiretroviral effect of IFN (as shown by the p24 antigen parameter) is more pronounced in patients with superior immune parameters.

AIDS-Related Complex

Analysis of polyomavirus enhancer-effect on DNA replication and early gene expression.

The polyomavirus enhancer is located adjacent to the origin of DNA replication and the transcriptional promoters. It has a cis-acting essential function in the initiation of both viral DNA synthesis and early transcription. The enhancer is activated by the binding of protein factors to specific sites in DNA. Mutants with deletions of the A- or the B-segment of the enhancer were constructed. In mouse 3T6 cells, the transcription of the viral early region was significantly decreased by deletion of the A-segment, but not by deletion of the B-segment. In contrast, the two deletions had a similar, moderately negative effect on viral DNA synthesis. However, the presence of DNA with a wild-type enhancer in doubly transfected cells resulted in very strong interference with the replication, but not with the transcription, of deletion mutant DNA. DNA of the deletion mutants were subjected to site-directed mutagenesis of the remaining enhancer segment. Three non-viable mutants were isolated. All three had base-pair changes in the A-segment affecting immediately adjacent binding sites of cellular protein factors. The mutants had lost the enhancer activity on the early promoter, but only one of them with multiple base substitutions had lost the capacity of DNA replication. Together, the results suggest that different aspects of enhancer function determine the activity in initiation of transcription and replication.

Amino Acid Sequence

Analogues of parathyroid hormone modified at positions 3 and 6. Effects on receptor binding and activation of adenylyl cyclase in kidney and bone.

Predictive and spectroscopic methods were used to develop a model of the structures of the 1-34 peptides of parathyroid hormone (PTH) and the PTH-related protein (PTHrP). Circular dichroism (CD) studies of bovine PTH-(1-34) and human PTHrP-(1-34)amide in the presence of trifluoroethanol suggest the presence of 24-26 alpha-helical residues. For both peptides, interactions between amino- and carboxyl-region alpha-helices are predicted to result in a hydrophobic core with externally facing hydrophilic residues that include probable determinants of receptor binding and activation. Two such residues, Ser3 and Gln6, are conserved in all known members of the PTH/PTHrP family. We have synthesized 13 novel analogues of bovine PTH-(1-34) monosubstituted at positions 3 and 6 and have determined their biological activities in renal and bone cell radioreceptor and adenylyl cyclase assays. Position 3 analogues displayed biological activity that was reduced in direct proportion to the volume of the substituent side-chain. Position 6 analogues also displayed reduced biological activity, but no simple correlation with side-chain volume or hydrophobicity was evident. The analogues fully displaced labeled PTH from binding sites in renal membranes and bone cells, but [Phe3]bPTH-(1-34), [Tyr3]bPTH-(1-34), [Phe6] bPTH-(1-34), and [Ser6]bPTH-(1-34) were only partial agonists in one or both adenylyl cyclase assays. Of these, [Phe3]bPTH-(1-34) and [Phe6]bPTH-(1-34) were tested for antagonist activity and were found to inhibit the activation of adenylyl cyclase in response to bPTH-(1-34) or hPTHrP-(1-34)amide. These results indicate that positions 3 and 6 contribute important determinants of PTH receptor binding and activation. Modification at these positions represents a novel approach to the development of antagonists of PTH action.

Adenylyl Cyclases

Post-antibiotic effect of beta-lactam antibiotics on gram-negative bacteria in relation to morphology, initial killing and MIC.

The in vitro post-antibiotic effect (PAE) of cefepime, cefotaxime, ceftazidime and imipenem on reference strains of Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa and Serratia marcescens were evaluated by bioluminescence assay of bacterial ATP. In parallel with the PAE determination, initial killing and morphology studies were performed. Imipenem produced greater than 1 h PAE on all strains tested, cefepime and cefotaxime on four strains and ceftazidime only on one of the strains tested. The length of the PAE on different strains did not correlate in the same way to MIC. Imipenem induced greater than 1 h PAE at 1/4-2 MIC while the cephalosporins caused greater than 1 h PAE at 4-256 x MIC. A PAE exceeding 1.2h was seen concomitantly with spheroplasts but there was not necessarily strong (greater than or equal to 99%) initial killing at the same time. The PAE duration at greater than or equal to 99% initial killing varied between 2.0 h and 5.0 h. When the cephalosporins produced less than 1 h PAEs, this was seen concomitantly with production on filaments and weak initial killing. The bioluminescence method was not jeopardized by filament formation and no negative PAE was found in contrast to the viable count method. The study showed that neither a certain multiple of MIC, the presence of spheroplasts nor strong initial killing can predict the length of PAE for beta-lactam antibiotics on gram-negative bacteria.

Anti-Bacterial Agents