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J K Andersen

Publications and source records attributed to J K Andersen.

At least 37 records · Page 2Linked to original sources

Variations in virulence between different electrophoretic types of Listeria monocytogenes.

A total of 245 strains of Listeria monocytogenes, representing 33 different electrophoretic types (ETs), were examined quantitatively for haemolytic activity. No significant difference was observed in the mean haemolytic activity between different ETs. Eighty four out of 91 strains examined were found to be virulent for chick embryos. Strains belonging to ET 2 and ET 4 were found to be less virulent than strains of other ETs (P = 0.0447). Furthermore, strains from clinical cases were found to be more virulent (P = 0.0002) than strains from foods (the MTD among clinical strains was 2.46 in mean compared with 3.64 among food isolates). The explanation for this may be that more virulent strains are more prone to cause human infection. It is, however, also possible that strains of L. monocytogenes may become more virulent while multiplying in a living organism compared with multiplying in foods.

Animals↗

Incidence and control of Listeria monocytogenes in foods in Denmark.

The Danish regulatory policy on Listeria monocytogenes in foods is based on the principles of HACCP and was developed using a health risk assessment approach. The Danish policy focuses examinations and criteria for L. monocytogenes in ready-to-eat foods and is based on a combination of inspection and product-testing. Based on current epidemiological information from several countries, a concentration of L. monocytogenes not exceeding 100 cfu/g of food at the time of consumption, seems to be of low risk to the consumers. In Denmark, ready-to-eat foods have been placed into six categories where absence of L. monocytogenes in 25 g is required in foods heat treated in the final package and in heat-treated as well as preserved, non heat-treated foods which can support growth within the shelf life. This level is necessary in foods capable of supporting growth, in order not to exceed 100 L. monocytogenes per g at the point of consumption. In heat-treated and preserved foods, which are not supportive of growth within the shelf-life and for raw, ready to eat foods, a level below 10 L. monocytogenes per g is regarded acceptable. A level between 10 and 100 L. monocytogenes per g is not satisfactory and a level above 100/g is not acceptable. Data on the qualitative and quantitative occurrence of L. monocytogenes in foods in Denmark are presented and discussed. In 1997 and 1998, greater than 15,000 samples from different categories of food were examined (semi-quantitatively) for the presence of L. monocytogenes. A significant difference could be seen in the number of samples containing more than 100 L. monocytogenes per g, between different categories of foods (1997, P = 0.001; 1998, P = 0.016). In 1997, preserved meat products and preserved fish products and to a lesser extent vegetables and meat or vegetable mayonnaise were more likely to contain high numbers (i.e. above 100 cfu/g) of L. monocytogenes than other food categories. In 1998, preserved meat products, but also heat-treated meat products, vegetables and meat or vegetable mayonnaise had the highest frequency of samples with > 100 L. monocytogenes per g. In a survey performed in 1994 and 1995, 1.3% of ready-to-eat food samples (heat-treated meat products, preserved meat and fish products) were found to be contaminated with L. monocytogenes at a level above 100 cfu/g. The samples included in this survey were primarily products produced by authorized companies and were comprised mainly of vacuum packed products or products packed in modified atmosphere and with long shelf lives, typically above several weeks. The corresponding percentages of positive samples primarily processed in the retail outlets (heat-treated meat products, preserved meat and fish products) in 1997 and 1998 were 0.3% and 0.6%, respectively. The results suggest that ready-to-eat meat and fish products with extended shelf-lives produced by authorized companies are more likely to contain high numbers (> 100 cfu/g) of L. monocytogenes than products processed in the retail sector which often have a shorter shelf life.

Animals↗

Brain gamma-glutamyl cysteine synthetase (GCS) mRNA expression patterns correlate with regional-specific enzyme activities and glutathione levels.

The first and rate-limiting reaction in the formation of glutathione is catalyzed by gamma-glutamylcysteine synthetase (GCS), a dimer composed of a catalytic heavy and a regulatory light subunit. We previously found that heavy subunit GCS mRNA appears to be expressed at high levels in the hippocampus, cerebellum, and cortex of murine brain and at lower levels in the neostriatum (Kang et al. [1997] NeuroReport 8:2053). Here we report that variations in expression of light subunit GCS mRNA in murine brain resembles that of the heavy subunit mRNA with a few minor exceptions. Moreover, levels of GCS activity and glutathione levels in various brain regions appear to correspond to levels of expression of both GCS mRNA subunits. Based on these data, differences in the distribution of expression of the GCS subunits in the brain may therefore have major implications for the susceptibility of various brain regions to oxidative stress and/or mitochondrial damage.

Animals↗

Establishment of long-term CD154-dependent porcine B-cell cultures.

Cells of the B-cell lineage play an essential part in the immune response, not only as the producers of antigen-specific antibodies, but also as antigen-presenting cells. Unlike T cells, however, the establishment of long-term normal B-cell lines has proved to be exceedingly difficult. In this paper we demonstrate that cell membrane-expressed CD154 (CD40 ligand) is able to support the continual growth of porcine mesenteric lymph node B-cell cultures for more than 4 months without the addition of exogenous cytokines, such as interleukin-4 (IL-4). Addition of IL-4, but not interferon-gamma (IFN-gamma) or IL-13, to these cultures enhanced proliferation, as, to a lesser extent, did addition of IL-2. Interestingly, however, whilst IFN-gamma-supplemented cultures largely consisted of immunoglobulin M (IgM)-positive cells, cultures with IL-13 or IL-4 contained a significantly increased proportion of IgG-positive cells.

Animals↗

Systematic characterization of porcine ileal Peyer's patch, I. apoptosis-sensitive immature B cells are the predominant cell type.

It is now apparent that the Peyer's patches of some species exhibit structural, functional and developmental heterogeneity. In sheep, for example, the ileal Peyer's patch (IPP) is the primary, antigen-independent site for the generation of the primary immunoglobulin repertoire and consequent production of the systemic B-cell pool. The pig has three distinct Peyer's patches, including an IPP, but the functional status of this organ, as primary or secondary lymphoid tissue, is not clear. Here, we have systematically characterized pig IPP follicular lymphocytes and show that about 90% B cells that are positive for surface immunoglobulin G (sIgM+) and express an immature phenotype characterized by expression of myeloid marker sWC3 (74-22-15) and two molecules recognized by IPP B-cell-specific monoclonal antibodies (F10/4, F12/35). Extensive apoptosis in vivo and in vitro was demonstrated by electron microscopy, immunohistology with TdT-mediated dUTP nick end labelling, DNA analysis and fluorescence-activated cell sorter analysis. Thus, when isolated IPP follicular cells were incubated at 37 degrees in vitro, the majority of them became apoptotic. The few that survived, however, had lost their expression of sWC3, F10/4, F12/35, but showed an increased expression of sIgM and major histocompatibility complex class II indicating that such surviving cells were of a more mature phenotype. Although more T cells were observed in porcine IPP follicles than in sheep IPP, CD3+ cells comprised less than 5% of the IPP follicular lymphocytes. Thus, the results clearly indicate that pig IPP is equivalent to sheep IPP.

Animals↗

Systematic characterization of porcine ileal Peyer's patch, II. A role for CD154 on T cells in the positive selection of immature porcine ileal Peyer's patch B cells.

We previously demonstrated that the majority (>/= 90%) of porcine ileal Peyer's patch (IPP) follicular cells are immature B cells destined to die by apoptosis, when incubated at 37 degrees. In this paper we approached the mechanisms responsible for positive selection of porcine IPP follicular immature B-cell selection, by screening for various cell types, cytokines and polyclonal and monoclonal antibodies for promoting the survival of IPP B cells. Of these reagents, only CD3 cross-linked purified T cells from mesenteric lymph nodes were able to rescue IPP follicular B cells from apoptosis, although polyclonal anti-IPP lymphocyte antibodies delayed apoptosis. This survival effect could be reproduced simply by incubating IPP follicular B cells with soluble and cell membrane-expressed CD154, an observation consistent with the demonstrated presence of CD40 and CD154 on porcine IPP follicular B cells and activated T cells, respectively. The IPP follicular B cells rescued in this manner expressed a more mature surface marker phenotype. Immunohistology and fluorescence-activated cell sorter analysis demonstrated that subpopulations of IPP follicular T cells (less than 0.5%) express CD154. Thus, perhaps unexpectedly, CD154 on T cells may play a role in the positive selection of immature B cells in the porcine IPP. The origin and control of the activated T cells identified within the porcine IPP remains to be investigated.

Animals↗

The role of iron in Parkinson disease and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity.

Parkinson disease (PD) involves the specific degeneration of dopaminergic neurons of the pars compacta of the substantia nigra. Although the cause of the degeneration of nigrostriatal dopaminergic neurons in PD is unknown, there is significant evidence to suggest that oxidative stress may be involved in this process. This review specifically examines the current status of evidence suggesting iron may contribute to oxidative damage associated with PD.

Animals↗

African swine fever virus: a B cell-mitogenic virus in vivo and in vitro.

The two major characteristics of pathogenesis in African swine fever virus (ASFV) infections of domestic pigs are massive B-cell apoptosis and haemorrhage. The effects of ASFV on porcine B cells have therefore been systematically examined in vivo, by using virus-infected pigs and SCID-Beige mice reconstituted with porcine bone marrow, and in vitro, by using porcine B-cell lines and B cells from normal and ASFV-infected pigs. Secretion of porcine Ig was stimulated by ASFV both in vivo and in bone marrow cultures in vitro, with the virulent Malawi isolate of ASFV being the most effective. Stimulation of Ig secretion in vitro depended on the presence of ASFV-infected macrophages and did not occur with supernatants from ASFV-infected macrophages. Although the virus alone did not stimulate proliferation of purified B cells in vitro, it was co-stimulatory with CD154 (CD40 ligand). The B cells recovered from ASFV-infected porcine lymphoid tissue were of activated surface marker phenotypes and, interestingly, expressed diminished levels of the B-cell co-stimulatory surface molecule CD21. In addition, they were highly sensitive to IL-4 and CD154. These results may be integrated into a model of pathogenesis in which those B cells activated indirectly as a result of virulent ASFV infection of macrophages are not rescued from apoptosis through interaction with CD154, due to the drastic depletion of T cells that occurs early in infection. The consequently diminished specific anti-ASFV antibody response would favour survival of the virus, with the non-specific hypergammaglobulinaemia being perhaps another example of pathogen-mediated immune deviation.

African Swine Fever↗

Use of genetically engineered mice as models for exploring the role of oxidative stress in neurodegenerative diseases.

A growing body of evidence has suggested that oxidative stress may play a major role in the degeneration of neurons associated with several neurological diseases of aging including ALS, Parkinson's, and Alzheimer's disease; this has been the topic of numerous previous reviews and opinion papers (e.g. 1-10). The ability to construct genetically engineered mouse lines containing targeted mutations has done much to aid in the assessment of the role of reactive oxygen species (ROS) in both the initiation as well as the progression of these diseases and has markedly advanced research in the field. Most importantly, the creation of genetic animal models has strengthened the argument that antioxidants may be a useful therapy in the treatment of these types of disorders.

Animals↗

Increased expression of monoamine oxidase-B results in enhanced neurite degeneration in methamphetamine-treated PC12 cells.

In vivo administration of methamphetamine (MA) produces selective damage to dopaminergic nerve terminals, which is hypothesized to be due to release of dopamine from synaptic vesicles within the terminals, allowing the generation of reactive oxygen species (ROS) via dopamine metabolism. Hydrogen peroxide formed during this reaction can interact with free iron to form hydroxyl radicals, which can oxidize proteins, nucleic acids, and membrane lipids, leading to terminal degeneration. Elevation of activity of the dopamine-metabolizing enzyme monoamine oxidase (MAO) in nerve growth factor-treated PC12 cells resulted in a substantial rise in products of dopamine metabolism following MA treatment, including 3,4-dihydroxyphenylacetic acid and hydroperoxides, as well as an increase in lipid peroxidation and a decrease in neurite number and length compared with control cells. These latter effects could be reversed by treatment with the MAO-B specific inhibitor, deprenyl. These data suggest that dopamine metabolism and subsequent ROS production may be key elements in MA-induced neurite degeneration in dopaminergic neurons.

3,4-Dihydroxyphenylacetic Acid↗

Cloning/brain localization of mouse glutamylcysteine synthetase heavy chain mRNA.

Glutathione (GSH) is considered the primary molecule responsible for peroxide removal from the brain. Inhibition of its rate-limiting synthetic enzyme, glutamylcysteine synthetase (GCS), results in morphological damage to both cortical and nigral neurons in rodents. Here, we report cloning of the catalytic heavy chain GCS mRNA from mouse and its localization in the murine brain. Heavy chain GCS appears to be localized in glial populations in the hippocampus, cerebellum and olfactory bulb, with lower levels of expression in the cortex and substantia nigra. Variations in GCS levels and subsequent GSH synthesis may explain differences in susceptibility to neuropathology associated with oxidative stress noted in these various brain regions.

Amino Acid Sequence↗

Elevated expression of glutathione peroxidase in PC12 cells results in protection against methamphetamine but not MPTP toxicity.

In vivo administration of either 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or methamphetamine (MA) produces damage to the dopaminergic nervous system which may be due in part to the generation of reactive oxygen species (ROS). The resistance of superoxide dismutase (SOD) over-expressing transgenic mice to the effects of both MPTP and MA suggests the involvement of superoxide in the resulting neurotoxicity of both compounds. Superoxide can be converted by SOD to hydrogen peroxide, which itself can cause cellular degeneration by reacting with free iron to produce highly reactive hydroxyl radicals resulting in damage to proteins, nucleic acids and membrane phospholipids. Hydrogen peroxide has also been reported to be produced via inhibition of NADH dehydrogenase by MPP + formed during oxidation of MPTP by MAO-B and by dopamine auto-oxidation following MA-induced dopamine release from synaptic vesicles within nerve terminals. To test whether hydrogen peroxide is an important factor in the toxicity of either of these two neurotoxins, we created clonal PC12 lines expressing elevated levels of the hydrogen peroxide-reducing enzyme glutathione peroxidase (GSHPx). Elevation of GSHPx levels in PC12 was found to diminish the rise in ROS levels and lipid peroxidation resulting from MA but not MPTP treatment. Elevated levels of GSHPx also appeared to prevent decreases in transport-mediated dopamine uptake produced via MA administration as well as to attenuate toxin-induced cell loss as measured by either MTT reduction or LDH release. Our data, therefore, suggest that hydrogen peroxide production likely contributes to MA toxicity in dopaminergic neurons.

Animals↗

Decreased glutathione results in calcium-mediated cell death in PC12.

Neuronal damage in certain cellular populations in the brain has been linked to oxidative stress accompanied by an elevation in intracellular calcium. Many questions remain about how such oxidative stress occurs and how it affects calcium homeostasis. Glutathione (GSH) is a major regulator of cellular redox status in the brain, and lowered GSH levels have been associated with dopaminergic cell loss in Parkinson's disease (PD). We found that transfection of antisense oligomers directed against glutamylcysteine synthetase (GCS), the rate-limiting enzyme in GSH synthesis, into PC12 cells resulted in decreased GSH and increased levels of ROS. Decreased GSH levels also correlated with an increase in intracellular calcium levels. Data from this study suggest that dopaminergic neurons are very sensitive to decreases in the internal oxidant buffering capacity of the cell caused by reductions in GSH levels, and that alterations in this parameter can result in disruption of calcium homeostasis and cell death. These results may be of particular significance for therapeutic treatment of PD, as those dopaminergic neurons that are spared in this disorder appear to contain the calcium binding protein, calbindin.

Animals↗

Genetic elevation of monoamine oxidase levels in dopaminergic PC12 cells results in increased free radical damage and sensitivity to MPTP.

Production of hydrogen peroxide as a by-product of the breakdown of catecholamines by the enzyme monoamine oxidase (MAO) has been hypothesized to contribute to the increased proclivity of dopaminergic neurons for oxidative injury. We established clonal dopaminergic PC12 cell lines which have elevated MAO activity levels resulting from transgenic expression of the B isoform of the enzyme. Both MAO-A and MAO-B have relatively equivalent affinities for dopamine, and since PC12 primarily express the A and not the B form of the enzyme, this allowed us to distinguish the transgenic MAO activity in these cells from endogenous using the MAO-B specific substrate PEA. Elevation of MAO activity levels in the MAO-B+ cells resulted in higher levels of both free radicals and free radical damage compared with controls. In addition, increased MAO-B levels within PC12 cells caused a dose-dependent increase in sensitivity to the toxin MPTP. Our data suggests that oxidation of catecholamines by MAO can contribute to free radical damage in catecholaminergic neurons and that the low MAO-B activity levels found endogenously in these cells likely accounts for their relative resistance to MPTP toxicity.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Effect of buthionine sulfoximine, a synthesis inhibitor of the antioxidant glutathione, on the murine nigrostriatal neurons.

This study analyzed the effects of acute systemic treatment with buthionine sulfoximine (BSO), a synthesis inhibitor of the antioxidant reduced glutathione (GSH), on dopaminergic neurons of the murine nigrostriatal pathway. Part 1 of the study established a dose-response curve and the temporal pattern of GSH loss and recovery in the substantia nigra and striatum following acute BSO treatment. Part 2 of the study determined the effect of acute BSO treatment on the morphology and biochemistry of nigrostriatal neurons. We found that decreases in GSH levels had profound morphological effects, including decreased catecholamine fluorescence per cell, increased levels of lipid peroxidation and lipofuscin accumulation, and increased numbers of dystrophic axons in dopaminergic neurons of the nigrostriatal pathway. However, no measurable effects were observed in biochemical levels of either dopamine or its metabolites. These changes mimic those that have been reported to occur in the nigrostriatal system of rodents with advancing age. Our data suggest that reduction of GSH via BSO treatment results in the same types of nigrostriatal degenerative effects that occur during the aging process and consequently is a good model system for examining the role of GSH in protecting this area of the brain against the harmful effects of age-related oxidative stress.

3,4-Dihydroxyphenylacetic Acid↗

Decreases in protective enzymes correlates with increased oxidative damage in the aging mouse brain.

We used several biochemical assays to evaluate age-related changes in antioxidant enzyme levels vs. free-radical damage in the murine brain. We found levels of several free-radical scavenging enzymes in the brains of 24-month-old C57B1 male mice vs. 12-month-old animals were decreased, including superoxide dismutase (SOD), catalase, and glutathione reductase (GSSG-Rd). In addition, we found concomitant increases in the levels of several forms of free-radical damage including sensitivity to lipid peroxidation as measured by the thiobarbituric acid test, protein oxidation as measured by glutamine synthetase (Gln Syn) activity, as well as increases in oxidized glutathione (GSSG) levels, a measure of oxidative stress. These data suggest that decreases in levels of enzymes which ordinarily protect neuronal cells against oxidative stress with age may be responsible for increased levels of free-radical damage in the murine brain, or that these enzymes themselves are susceptible to inactivation by free radical molecules which increase with age in the brain.

Aging↗