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Biomedical subjects

M Nilsson

Publications and source records attributed to M Nilsson.

At least 199 records · Page 11Linked to original sources
PubMed1993

Biosynthesis and morphogenesis of group C rotavirus in swine testicular cells.

Polypeptide synthesis and morphogenesis of a group C rotavirus (AmC-1) adapted to a continuous swine testicular cell line was examined. SDS-PAGE analysis of 35S methionine labeled infected cell lysates revealed 9 viral polypeptides (122, 98, 79, 78, 43, 41, 35, 24, and 20 kD). Viral polypeptide synthesis appeared to be maximal at 7-10h post infection. Purified group C virus grown in the presence of trypsin was found to contain seven structural polypeptides (122, 98, 79, 53, 43, 41, and 30 kD) by protein blotting and five polypeptides (98, 79, 78, 43, and 41 kD) by immunoprecipitation with a hyperimmune rabbit antisera. Tunicamycin treatment, Concanavalin A binding, protein blotting, endo-H treatment and 2,6H-mannose labeling suggested that group C rotavirus contains one structural glycoprotein (41 kD) with a corresponding precursor mol. wt. of 37 kD and one not previously identified nonstructural glycoprotein (24 kD) with a corresponding precursor mol. wt. of < or = 20 kD. Electron microscopy of infected swine testicular cells revealed an assembly process for group C rotavirus similar to group A, with single-shelled particles budding through the rough endoplasmic reticulum with concomitant acquisition of a transient membrane.

Animals↗
PubMed1993

Automated immunochemical binding assay (flow-ELISA) based on repeated use of an antibody column placed in a flow-injection system.

A computer-controlled immunological binding assay intended for monitoring concentrations of interesting metabolites during bioprocesses has been developed. The main focus has been on speed, reliability, reproducibility and operational stability of the assay. The assay is based on the repeated use of a preparation of immobilized antibodies. Stability in the regeneration step is a prerequisite and was achieved by intermittent recalibration of the system. Furthermore, before immobilizing the polyclonal antibodies, they were purified on immobilized antigen and eluted using the same procedure as used in the assay cycle. Antibody preparation could be used for more than 50 cycles and in some cases for several hundred cycles. Since the calibration curve is valid for the whole life-time of the antibody column, recalibration only involved the registration of the column capacity. Experiments have been performed concerning monitoring of chromatographic separations as well as adsorption experiments from complex mixtures.

Animals↗
PubMed1993

GABA induces Ca2+ transients in astrocytes.

By using the Ca(2+)-sensitive indictor Fura-2/AM, the cytosolic Ca2+ levels [Ca2+]i were measured in type 1 astrocytes in rat cortical astroglial primary cultures, after stimulation with GABA, muscimol (GABAA agonist), or baclofen (GABAB agonist). We report the first evidence that stimulation of both GABAA and GABAB receptors evokes Ca2+ transients in type I astrocytes. Two types of Ca2+ responses were seen: the single-phase curve, which was the most common, and the biphasic, which consisted of an initial rise that persisted at the maximal or submaximal level. Both types of Ca2+ responses appeared with some latency. The responses were obtained in astrocytes grown for 12-16 days in culture and the response frequencies for all three agonists were 18% of the total number of examined cells. However, when the astrocytes were grown in a mixed astroglial/neuronal culture the response frequencies for all three agonists increased to 35% of the total number of examined cells. In some cells, the responses after GABA stimulation were blocked to baseline levels after exposure to bicuculline (GABAA antagonist). In other cells, bicuculline only slightly reduced the GABA-evoked responses, and the addition of phaclofen (GABAB antagonist) did not potentiate this partial inhibition. However, the muscimol-evoked rises in [Ca2+]i were completely inhibited after exposure to bicuculline, while the responses after baclofen could only be partly blocked by phaclofen. GABA evoked rises in [Ca2+]i which alternatively were inhibited (mostly) or persisted in Ca(2+)-free buffer. The rises in [Ca2+]i persisted, but were reduced, in Ca(2+)-free buffer after stimulation with muscimol, but were inhibited after baclofen stimulation. The GABA uptake blockers guvacine, 4,5,6,7-tetrahydroisoxazolo(4,5-c)pyridin-3-ol and nipecotic acid were also able to reduce the GABA-evoked rises in [Ca2+]i. However, the L-type Ca2+ channel antagonist nifedipine failed to influence on the GABA-evoked Ca2+ transients. The results suggest that type 1 astrocytes in primary culture express GABA receptors which can elevate [Ca2+]i directly or indirectly via Ca2+ channels and/or via release from internal Ca2+ stores. The results also suggest that GABA can have intracellular Ca(2+)-mobilizing sites since the GABA-evoked responses were reduced after incubation with GABA uptake blockers.

Animals↗
PubMed1993

Kappa-opioid receptors on astrocytes stimulate L-type Ca2+ channels.

Cultured astrocytes from the cerebral cortex of the rat respond to opioid kappa-receptor stimulation with a substantial elevation of the cytoplasmic free calcium, visualized through the use of the fluorescent calcium indicator Fura-2. The stimulation of kappa-receptors with U-50488H increases the level of calcium through a dose-related stimulatory effect on the transmembrane calcium influx. The kappa-receptor stimulation was completely blocked by the selective kappa-receptor blocker nor-binaltorphimine. Furthermore, the transmembrane calcium influx was completely blocked by nifedipine, indicating the involvement of L-type channels. The presence of L-type channels was verified by stimulation of L-type channels with Bay K8644. The effects of Bay K8644 were completely blocked by nifedipine. L-type channel-coupled kappa-receptors on astrocytes might represent a novel mechanism contributing to the depressant action of opioids on synaptic transmission via decreasing the availability of extracellular calcium necessary for presynaptic transmitter release.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1993

Effects of imipenem on Escherichia coli studied using bioluminescence, viable counting and microscopy.

The effects of imipenem on the growth of Escherichia coli ATCC 25922 were studied using a bioluminescence assay of bacterial ATP, microscopy and viable counting in iso-osmotic Mueller-Hinton broth (MHB) and hypo-osmotic nutrient broth (NB). Imipenem showed a post-antibiotic effect (PAE) of > 2 h for E. coli in both MHB and NB after 2 h exposure to 1 and 8 mg/L of imipenem when determined by bioluminescence and microscopy. The intracellular ATP level increased after 2 h exposure of E. coli to 1 mg/L of imipenem in MHB. In this culture there was a predominance of spheroplasts. These spheroplasts were large and osmotically fragile and a 10 min treatment in water-diluted MHB (hypo-osmotic) prior to the assays lysed the large spheroplasts. This reduced the intracellular ATP level and shortened the PAE when determined by bioluminescence, and caused more rapid initial killing and a negative PAE when determined by viable counting. At 8 mg/L imipenem in MHB and at all concentrations in NB there was a predominance of rods and only a small number of spheroplasts which all disappeared when the cultures resumed logarithmic growth. In these cultures there was a significant initial decrease in intracellular ATP. This study showed reasonable agreement between microscopy and bioluminescence, which are direct methods, for determining the initial killing and PAE of imipenem on E. coli. More rapid initial killing and shorter or no PAEs, were in general, obtained in both MHB and NB when determined by viable counting. However, the effective regrowth time, defined as the time for the bacterial density to increase 1 log10 from the pre-exposure inoculum, was independent of the method used for measuring regrowth in both MHB and NB.

Adenosine Triphosphate↗
PubMed1993

Postantibiotic effect of aminoglycosides on staphylococci.

The postantibiotic effects (PAEs) of amikacin, gentamicin, netilmicin and tobramycin on Staphylococcus aureus and S. epidermidis were determined in vitro by a bioluminescence assay of bacterial ATP. Five strains of S. aureus and two strains of S. epidermidis were exposed for 1 h to varying concentrations of these aminoglycosides. Following removal of the antibiotics by dilution, bacterial regrowth was monitored at hourly intervals. The duration of the PAE increased with increasing aminoglycoside concentration. The mean PAEs for the five S. aureus strains ranged from 5-10 h at clinically achievable aminoglycoside concentrations (16-32 mg/L of amikacin and 4-8 mg/L of gentamicin, netilmicin and tobramycin). The results for one of the strains of S. epidermidis were similar to those observed for the S. aureus strains, while the PAEs on the other less susceptible S. epidermidis strain were shorter (0.5-2.5 h). For comparison, two of the S. aureus strains were exposed for 1 and 2 h to a range of concentrations of dicloxacillin (0.25-32 mg/L); this agent induced a much shorter PAE (0-2.3 h). It may be important to take account of the PAE when designing dosing regimens.

Amikacin↗
PubMed1992

[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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗
PubMed1992

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↗