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Biomedical subjects

K Sandberg

Publications and source records attributed to K Sandberg.

At least 91 records · Page 5Linked to original sources
PubMed1989

Association between serum esterase (Es) type and starting proportion in Swedish Trotters: further observations.

A study was carried out to attempt to explain the basis of the association between the Es locus and starting proportion in Swedish Trotters which had been observed previously. The effect of Es genotype on starting proportion has diminished among horses born in the late 1970s. There are indications that the incidence of leg lesions varies between Es genotypes. If this is true, it is possible that the decreasing effect of the Es locus on starting proportion is due to the environmental changes which have been made at race tracks in the early 1980s to reduce the strain on the legs of the trotters.

Animals↗
PubMed1989

Characterization of the blood mononuclear leucocytes producing alpha interferon after stimulation with herpes simplex virus in vitro, by means of combined immunohistochemical staining and in situ RNA-RNA hybridization.

A procedure is described for combined immunohistochemical staining and in situ RNA-RNA hybridization of human peripheral blood mononuclear leucocytes (PBMC). These cells were first stimulated in vitro by herpes simplex virus (HSV)-infected fibroblasts and after 6 h fixed and stained with a panel of antibodies against differentiation antigens. Alpha interferon (IFN-alpha)-producing cells (IPC) were identified by in situ hybridization by means of a 35S-labelled IFN-alpha 2 cRNA probe. The IPC were infrequent, one in 200-5000 PBMC, but heavily labelled with the cRNA probe. They lacked antigens typical of T and B lymphocytes, and were also essentially negative for the Leu-M5 antigen, present on a majority of monocytes. However, 50% of IPC expressed OKM5 antigens, corresponding to the thrombospondin receptor. The IPC lacked the antigens present on null lymphocytes detected by OKT16, but most of them expressed HLA-DR, -DP and -DQ antigens. The IPC may represent a small subpopulation in the monocyte/macrophage lineage, resembling cells described as antigen presenting and stimulators of autologous mixed lymphocyte reactions. Alternatively, they constitute a subpopulation among the null lymphocytes.

Antibodies, Monoclonal↗
PubMed1989

A role for cGMP during tetanus toxin blockade of acetylcholine release in the rat pheochromocytoma (PC12) cell line.

In order to identify the specific molecular mechanisms involved in neurosecretion, we investigated the mechanism of action of tetanus toxin, a potent presynaptic neurotoxin, in the rat adrenal pheochromocytoma PC12 cell line. It has recently been reported that tetanus toxin is a potent inhibitor of the release of depolarization-evoked 3H-acetylcholine (ACh) from nerve growth factor-differentiated PC12 cells (Sandberg et al., 1989a). In PC12 cells, as in many neural tissue preparations, cGMP accumulation in intact cells increased 6- to 17-fold when stimulated with veratridine (200 microM), carbachol (1 mM), Ba2+ (2 mM), or K+ (30 mM). Preincubation of the cells with tetanus toxin inhibits this accumulation by greater than 95%. The toxin dose-inhibition curves for 3H-ACh release and cGMP accumulation are similar, with half-maximal doses of tetanus toxin seen at approximately 5 nM. The time courses for the development of the effects of tetanus on 3H-ACh release and on cGMP accumulation were also similar. Protocols which elevated intracellular cGMP levels reversed the action of the toxin. For example, evoked ACh release was restored in intoxicated PC12 cells by a 15 min exposure to 100 microM 8-bromo-cGMP. The half-maximal dose was observed at 50 microM nucleotide. Examination of the nucleotide specificity revealed that only cyclic guanine analogs were effective in reversing the effects of tetanus toxin. These results suggested that the inhibition of depolarization-evoked cGMP accumulation is causally related to the action of tetanus toxin on neurosecretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1988

Calcium mobilization by angiotensin II and neurotransmitter receptors expressed in Xenopus laevis oocytes.

Specific receptors for angiotensin II (AII) were expressed in albino Xenopus laevis oocytes co-injected with poly(A)+ mRNA isolated from rat adrenal cortex and the calcium-specific photoprotein, aequorin. In such oocytes, AII elicited rapid, dose-dependent rises in cytosolic free calcium with light emission responses up to 100-fold above basal levels. Ligand-induced light emission was also observed in oocytes injected with rat brain mRNA and stimulated with acetylcholine and glutamate. These findings demonstrate that mammalian AII receptors expressed in Xenopus oocytes are functionally linked to intracellular Ca2+ mobilization, and indicate the potential value of aequorin-injected oocytes for rapid and specific screening of mRNAs transcribed from expression libraries containing cloned receptor cDNAs.

Acetylcholine↗
PubMed1988

Complex ganglioside expression and tetanus toxin binding by PC12 pheochromocytoma cells.

Ganglioside expression and tetanus toxin binding were studied in the rat pheochromocytoma cell line PC12. Seven ganglioside species were readily detected in extracts of PC12 cells; two were identified as tri- and tetrasialogangliosides, which are common brain constituents but unusual components of neuronal cell lines. Carbohydrate composition, acid and enzyme hydrolyses, and mass spectral analysis revealed that the major species is GT 1b, a predominant mammalian brain ganglioside previously reported to support high affinity tetanus toxin binding (Rogers, T. B., and Snyder, S. H. (1981) J. Biol. Chem. 256, 2402-2407). Direct binding of 125I-tetanus toxin to PC12 gangliosides on TLC plates revealed selective binding to the tri- and tetrasialogangliosides. Radioiodinated toxin also bound with high affinity to intact PC12 cells or their isolated membranes. The binding affinity (Kd = 1.25 nM), density of receptors (Bmax = 238 pmol/mg of membrane protein), and dependence on pH, ionic strength, and temperature were similar to those previously reported for toxin binding to rat brain synaptic membranes. Differentiation of PC12 cells caused an increase in expression of the tri- and tetrasialogangliosides and a closely matched increase in tetanus toxin binding to cell membranes. These data provide evidence that complex gangliosides may act as tetanus toxin receptors, and demonstrate the utility of the PC12 cell line for studies of tetanus toxicity and complex ganglioside expression.

Adrenal Gland Neoplasms↗
PubMed1988

Determination of herpes simplex virus-induced alpha interferon-secreting human blood leucocytes by a filter immuno-plaque assay.

A filter immuno-plaque assay was developed which detects alpha interferon (INF-alpha)-secreting human peripheral blood leucocytes (PBL). Polyclonal anti-IFN-alpha antibodies were fixed to the nitrocellulose membrane bottoms of 96-well Millititer plates, which also contained monolayers of glutaraldehyde-fixed human WISH amnion cells infected by Herpes simplex virus type 1 (HSV). Such cells are potent IFN inducers and during a 16 h cocultivation with PBL, IFN-alpha was absorbed by the membrane-bound polyclonal antibodies around IFN-alpha-secreting cells. This IFN-alpha was detected with murine monoclonal antibodies against IFN-alpha and peroxidase-labelled antibodies against murine immunoglobulin, using diaminobenzidine as substrate. Distinctly stained plaques were seen, the frequency of which gave a minimal estimate of approximately 10 IFN-alpha-producing cells in 10(4) PBL (range in 12 blood donors 2.95-25.1). Fewer plaques than expected were seen at low PBL numbers per culture, one explanation being that cell interactions then limit the IFN-alpha response. The immuno-plaque assay should be useful in further studies of the cellular basis of the IFN-alpha response.

Enzyme-Linked Immunosorbent Assay↗
PubMed1987

Efficiency of ventilation in neonatal pulmonary maladaptation.

Lung physiology was studied in sixteen infants with pulmonary maladaptation (PMA) during the course of the disease and after clinical recovery. A sensitive nitrogen washout method was used. During the disease the infants showed reduced ventilatory efficiency and increased dead space. Total ventilation increased simultaneously, while alveolar ventilation was maintained. The majority of the infants showed greater functional residual capacity during the disease than after clinical recovery. The results suggest that gas mixing efficiency is impaired in infants with PMA and that this might be due to effects on the small airway function in the lungs.

Humans↗
PubMed1987

Pulmonary response to group B streptococcal toxin in young lambs.

Marked respiratory distress is seen in severe early onset group B beta-hemolytic streptococcal (GBS) disease in newborn infants. To investigate the pathophysiological effects of a polysaccharide toxin from GBS type III cultures, obtained from an infant who died from this disease, young chronically instrumented, unanesthetized lambs were studied with measurements of lung mechanics, lung volumes, ventilation, hemodynamics, and lung vascular permeability. Intravenously administered GBS toxin resulted in a biphasic response with an early threefold increase in total lung resistance, 40% decrease in dynamic lung compliance, and 30% increase in minute ventilation coinciding with hypoxemia, pulmonary hypertension, and fever. A second phase of the response followed consisting of less prominent changes in these variables as well as increased lung lymph protein clearance compatible with increased vascular permeability. The temporal close relationship between marked leukopenia and increased lung lymph thromboxane B2 concentrations to the simultaneously occurring pulmonary hypertension and changes in lung mechanics suggests that leukocytes and thromboxane A2 may be mediators of these GBS toxin-induced effects.

Airway Resistance↗
PubMed1987

The role of granulocytes in the pulmonary response to group B streptococcal toxin in young lambs.

Marked leukopenia and sequestration of granulocytes in the lung are consistently seen in severe early onset group B streptococcal (GBS) disease in human infants. To investigate the role of granulocytes as potential mediators in the pulmonary pathophysiology of this disease, the effects of intravenously administered GBS type III toxin were studied in young lambs before and after granulocyte depletion with hydroxyurea. Granulocyte depletion markedly reduced the 4-fold increase in total lung resistance and the decrease in dynamic compliance observed after GBS toxin. Granulocyte depletion significantly attenuated the pulmonary hypertension, hypoxemia and increased minute ventilation present during the first phase of the response (0.5-1 h after GBS toxin). It did not significantly alter the increase in body temperature, the marked increase in lung lymph thromboxane B2 concentrations during the first phase or the increase in lung lymph flow and protein clearance during the second phase of the response (3.5-5 h after GBS toxin). The results indicate that granulocytes are involved as mediators of the changes in lung mechanics seen after GBS toxin infusion in young lambs. Granulocytes contribute to the pulmonary hypertension and decrease in arterial oxygenation, but other mediators appear to be responsible for the injury of the vascular endothelium.

Animals↗
PubMed1987

Lung function in newborn infants with tachypnea of unknown cause.

Twenty-one infants with tachypnea (f greater than 60/min) lasting more than 2 h and diagnosed as mild respiratory disease or pulmonary maladaptation according to previously presented criteria were studied during the course of the disease and after clinical recovery. Lung physiology (total and alveolar ventilation, efficiency and distribution of ventilation, functional residual capacity, and lung mechanics) was studied in combination with clinical data. The pathophysiological findings were characterized by increased total ventilation but normal alveolar ventilation, reduced efficiency of ventilation but more even distribution of ventilation (nitrogen elimination pattern) during disease than after clinical recovery, hyperinflation, reduced dynamic lung compliance but unaffected specific lung conductance. Infants with low gestational ages were most severely affected and had longer duration of disease than full-term infants. Our findings suggest that this condition is caused by small airway disease. Disturbances in normal pulmonary adaptation with abnormal retention of lung fluid is the most probable cause.

Functional Residual Capacity↗
PubMed1986

Effects of delivery by caesarean section on lung function in healthy newborn infants.

With the aim of extending previous studies showing differences in lung function after birth between infants delivered vaginally (VD) and by Caesarean section (CS) we investigated lung volumes, ventilation, efficiency of ventilation, and lung mechanics in 24 healthy, full term infants with no clinical signs of respiratory disease, 12 after VD and 12 after CS. Measurements were made on two occasions: 2 and 26 hours after birth. At 2 hours no differences in any measured quantity were found between the groups. The only difference found 24 hours later was that the average thoracic gas volume (TGV), was lower in infants after CS than after VD. The difference in functional residual capacity was, however, not significant. This means that the difference in TGV, previously also found by other workers, did not affect the ventilated air space. Our results do not support the theory of general inferiority in lung performance after birth in healthy, full term infants without respiratory disease delivered by CS.

Cesarean Section↗
PubMed1986

Characterization of the binding and internalization of tetanus toxin in a neuroblastoma hybrid cell line.

Tetanus toxin is known to bind neuronal tissue selectively. To study the interactions of this potent neurotoxin in an intact cell system, the binding of 125I-tetanus toxin was characterized in a neuroblastoma retina hybrid cell line, N18-RE-105. The binding of 125I-tetanus toxin to membranes prepared from N18-RE-105 cells showed many similarities to the interactions of 125I-toxin with rat synaptic membranes. The binding was decreased with increasing temperature, ionic strength, and pH. 125I-Toxin bound to membranes with high affinity: KD = 0.62 +/- 0.05 nM; Bmax = 196 +/- 45 pmol/mg protein. Quantitative thin-layer chromatography and acid-degradation analysis revealed that N18-RE-105 cells contained polysialogangliosides GD1a and GT1b in high concentrations. An assay was developed to quantitate surface-bound and internalized 125I-tetanus toxin by exploiting the observation that surface-bound 125I-toxin is susceptible to pronase digestion. When cells were incubated with 125I-tetanus toxin at 0 degree C, all of the bound 125I-toxin could be degraded with pronase. In contrast, when the incubations were performed at 37 degrees C, within 10 min about 50% of the total cell-associated 125I-toxin was pronase-resistant. Temperature pulse experiments demonstrated that 125I-tetanus toxin that was bound to cells at 0 degree C rapidly disappeared from the surface when the cells were warmed to 37 degrees C, as revealed by the appearance of pronase-resistant radioactivity. This internalization was sensitive to metabolic inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Characterization of [3H]hemicholinium-3 binding associated with neuronal choline uptake sites in rat brain membranes.

Hemicholinium-3 (HCh-3) is a potent and specific inhibitor of the high-affinity choline transport process (HAChT) localized on cholinergic neurons. In this study, the specific binding of [3H]HCh-3 (120 Ci/mmol) was characterized in crude synaptic membranes prepared from rat brain. The binding of [3H]HCh-3 to forebrain membranes was saturable, reversible and specific with an apparent Kd under optimal conditions of 35 nM and a Bmax of 56 fmol/mg protein. The potency of various HAChT inhibitors correlated with their apparent affinities for the specific [3H]HCh-3 binding site. The specific binding of [3H]HCh-3 exhibited an uneven regional distribution in the adult rat brain that corresponded to the activity of the HAChT in these regions. Transsection of the fornix, which causes a degeneration of the septal hippocampal cholinergic pathway, resulted in comparable reductions of the specific [3H]HCh-3 binding and the specific activity of choline acetyltransferase, a presynaptic marker for cholinergic terminals in the hippocampal formation; the lesion did not affect the specific activity of glutamic acid decarboxylase, a presynaptic marker for GABAergic neurons within the hippocampus. Maximal binding occurred in the presence of 200 mM NaCl: potassium, lithium, rubidium and calcium substituted poorly for sodium; and bromide, fluoride, iodide, sulfate and phosphate were less effective anions than chloride. Increasing concentrations of NaCl increased the affinity of the site for [3H]HCh-3 with no significant effect on the maximal number of sites; the enhancement of affinity was due to a selective slowing of the rate of dissociation of the ligand from its binding site. These findings indicate that [3H]HCh-3 binds to the carrier site mediating the HAChT on cholinergic neurons; thus, this radioligand may be a useful probe for investigating this presynaptic component (HAChT) of cholinergic neurons.

Animals↗
PubMed1985

Method for the quantitation and characterization of the cholinergic neurotoxin, monoethylcholine mustard aziridinium ion (AF64A).

Monoethylcholine aziridinium ion (AF64A), which is generated from the precursor, acetylcholine mustard hydrochloride, exerts selective neurotoxic effects against brain cholinergic neurons when injected intracerebrally. Conditions associated with optimal generation of AF64A were examined. The results indicate that hydrolysis and cyclization of the precursor are optimal at 25 degrees C with the former occurring at pH 9.0 and the latter at pH 7.3. The aziridinium ion is best stabilized at pH 7.3 at 4 degrees C.

Aziridines↗
PubMed1985

AF64A: an active site directed irreversible inhibitor of choline acetyltransferase.

Ethylcholine mustard aziridinium ion (AF64A, MEChMAz) has been proposed as a cholinergic neuron-specific neurotoxin. We report that in further studies on its mechanism of action incubation of the cholinergic neuroblastoma X glioma cell line, NG-108-15, with 100 microM AF64A resulted in a rapid decrease in cellular choline acetyltransferase (ChAT) activity which preceded cytotoxicity. Thus, a 60-85% decrease in ChAT activity was measured within 5 h of AF64A exposure, whereas cell lysis (measured as the release of the cytosolic enzyme lactate dehydrogenase into the medium) did not become apparent until 18 h of AF64A exposure. This led us to examine the effects of AF64A on partially purified ChAT. We report a concentration- and time-dependent inhibition of partially purified ChAT by AF64A that could not be reversed by dialysis but could be prevented by coincubation of the enzyme and AF64A with choline but not with acetyl-coenzyme A. We present kinetic evidence that choline and AF64A compete for the same site on the enzyme. In addition, thiosulfate, which inactivates the aziridinium ion, eliminated AF64A's capacity to inhibit the enzyme. AF64A also irreversibly inhibited partially purified choline kinase and acetylcholinesterase but not lactate dehydrogenase, alcohol dehydrogenase, carboxypeptidase A, or chymotrypsinogen, enzymes that do not use choline as a substrate or product. Thus, the data suggest that AF64A acts as an irreversible active site directed inhibitor of ChAT and possibly other enzymes recognizing choline.

Animals↗
PubMed1984

Effects of intrastriatal injections of the cholinergic neurotoxin AF64A on spontaneous nocturnal locomotor behavior in the rat.

Rats with bilateral striatal cholinergic lesions induced by the neurotoxin AF64A were found to have increased spontaneous nocturnal locomotor activity as measured in terms of the amount of time spent moving, the total distance traveled and the number of episodes and time of stereotypic behaviors. No significant differences between control and AF64A injected rats were found in the actual velocities between the two groups indicating that the rats were simply spending more time moving rather than moving with greater speed. Striatal activity of choline acetyltransferase (CAT) was significantly decreased in AF64A treated rats compared to controls, whereas the activities of glutamate decarboxylase (GAD) were not. Furthermore, there were no significant differences between groups in enzymatic activities of CAT and GAD in either the cortex or the hippocampus, indicating that the lesion was restricted to the striatum. The hyperactivity found in these rats after intrastriatal injection of AF64A supports a role for the striatal cholinergic system in locomotor behavior.

Acetylcholine↗
PubMed1984

Selective cholinergic neurotoxin: AF64A's effects in rat striatum.

The selective neurotoxic effects of the aziridinium ion of ethylcholine (AF64A) have been examined after stereotaxic injection into the rat striatum. In a dose-response study (2-26 nmol), 8 nmol caused a 46% decrease in striatal choline acetyltransferase (CAT) activity with minimal effects on the activities of glutamate decarboxylase (GAD) and tyrosine hydroxylase (TH) at 7 days. Maximal CAT reductions of 78-82% occurred with doses of 16-26 nmol which also caused dose-related decreases in GAD and TH activities that paralleled the progressive decrements in CAT. A time course study with 8 nmol indicated a rapid 20% reduction of CAT activity by 12 h and an additional gradual fall of 20% over the next week; TH and GAD activities were not significantly reduced. The selective inhibition of CAT activity persisted for at least 3 months. Histological examination of Nissl stained sections revealed an area of nonspecific damage at the injection site with an abrupt border surrounded by apparently normal striatal neuropil; however; neuronal perikarya staining intensely for acetylcholinesterase were not reduced. These preliminary findings strongly suggest that AF64A has selective neurotoxic effects against striatal cholinergic neurons while relatively sparing striatal GABAergic intrinsic neurons or dopaminergic afferents.

Animals↗