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Biomedical subjects

A Karlsson

Publications and source records attributed to A Karlsson.

At least 73 records · Page 4Linked to original sources
PubMed2000

Phorbol myristate acetate induces neutrophil NADPH-oxidase activity by two separate signal transduction pathways: dependent or independent of phosphatidylinositol 3-kinase.

The neutrophil NADPH-oxidase can be activated by protein kinase C (PKC) agonists such as phorbol myristate acetate (PMA), resulting in superoxide anion release. This superoxide release is independent of phosphatidylinositol 3-kinase (PI 3-kinase) because the inhibitor wortmannin does not affect the response. In this study, PMA is shown to also induce a wortmannin-sensitive NADPH-oxidase activation, however, not resulting in release of superoxide but in intracellular production of the radical. This indicates that two pools of NADPH-oxidase, one localized in the plasma membrane and the other in the granule membranes, are separately regulated and the signal transduction pathways leading to activation of these pools differ regarding involvement of PI 3-kinase. Activation of both pools was dependent on ERK/MAPK kinase (MEK) activity and protein phosphatase 1 and/or 2A. As the two oxidase responses were differently affected by the inhibitor Gö-6850, different PKC isozymes are suggested to take part in the two signal transduction pathways.

Androstadienes↗
PubMed2000

Amplitude and frequency analysis of force plate data in sitting children with and without MMC.

OBJECTIVE: To investigate if force plate measurements can be used to detect postural sway differences in sitting children with and without myelomeningocele (spina bifida).BACKGROUND. The postural sway has not been investigated in children with myelomeningocele previously. Since many of these children are not able to stand independently, force plate measurements during sitting could be one way to detect differences in their postural sway compared to normal children. However, there is very little published regarding assessment of seated postural sway.METHODS. Force plate measurements on 15 six years old children with myelomeningocele and 20 age-matched normal children were analysed. The standard deviation and the median frequency of the horizontal ground reaction force were used to characterise the body sway.RESULTS. The standard deviation of the force was larger only for some of the children with myelomeningocele as compared to the control group. The median frequency was significantly lower in the myelomeningocele group as compared to the control group. Visual input and seat base inclination did not influence the postural sway significantly. CONCLUSIONS: The results show that frequency analysis can be used to detect fundamental differences in postural sway that can not be observed visually. RelevanceIn this paper a new method for analysis of seated postural sway is described. The lack of relatively high spectral frequencies for children with myelomeningocele shows that the output from the postural control system differs as compared to the control group.

Acceleration↗
PubMed2000

Correlations between force plate measures for assessment of balance.

OBJECTIVE: To compare different force plate measures for assessment of postural stability during normal standing. Five types of measures were analysed: (1) The standard deviation of the horizontal ground reaction force, (2) the standard deviation of the centre of pressure, (3) the mean velocity of the centre of pressure, (4) movement strategy measures, and (5) the standard deviation of the vertical ground reaction force. BACKGROUND: Force plate measures such as types 1-3 are frequently used, but they seem to have limited usefulness for assessment of balance on an individual level. Hence, there is a need to investigate alternative force plate measures (such as types 4 and 5). METHODS: Force plate measures for 20 stroke patients were (a) compared to each other, and (b) compared to a clinical balance test, called Berg's balance test. RESULTS: There was generally a significant correlation (P<0.01) between measures of types 1-3. The movement strategy measures were not significantly correlated to any of the other measures. The vertical force measure was only significantly correlated to the antero-posterior mean velocity of the centre of pressure. There was a significant correlation between (a) Berg's balance test and the vertical force measure (r(s)=-0.76), and (b) the static part of Berg's balance test and the antero-posterior mean velocity of the centre of pressure (r(s)=-0.6). CONCLUSIONS: Measures of type 1, 2, 4 and 5 seem to quantify different aspects of human standing. The results also indicate that Berg's balance test and the vertical force measure quantify a similar aspect of the postural stability. RELEVANCE: The results from this study may facilitate the understanding and interpretation of force plate measures.

Adult↗
PubMed2000

Identification and characterization of SarH1, a new global regulator of virulence gene expression in Staphylococcus aureus.

The global regulators agr (accessory gene regulator) and sarA (staphylococcal accessory regulator) have been reported to be both activators and repressors of virulence gene expression in Staphylococcus aureus. How the effector of the agr system, RNAIII, interacts with target gene promoters is unknown. SarA, on the other hand, is a DNA-binding protein, which binds to conserved DNA motifs immediately upstream of both positively and negatively regulated promoters. Here, we searched for additional regulators that could explain the differential effects of RNAIII and SarA. Four differently regulated genes (hla, alpha-toxin; hld, RNAIII; spa, protein A; ssp, serine protease) were analysed for binding of potential regulatory proteins to the corresponding promoter DNA fragments, linked to magnetic beads. One protein (29 kDa), with affinity for all four promoters, showed a high degree of similarity to SarA and was named SarH1 (Sar homologue 1). Expression of sarH1 was strongly repressed by sarA and agr. Analysis of hla, hld, ssp and spa mRNAs in sarH1, sarA and agr mutants, and in sarA/sarH1 and agr/sarH1 double mutants, revealed that sarH1 has a strong repressive effect on hla and an activating effect on spa transcription. SDS-PAGE analysis of secreted proteins from the different mutants showed that the production of several other exoproteins was affected by sarH1. In conclusion, we show that both the agr-dependent suppression of protein A production and the sarA-dependent stimulation of alpha-toxin production is mediated via a new regulator, SarH1, which belongs to a family of Sar homologues.

Amino Acid Sequence↗
PubMed2000

The multifunctional deoxynucleoside kinase of insect cells is a target for the development of new insecticides.

The antiherpetic agent (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) was found to be an efficient substrate for recombinant Drosophila melanogaster-deoxyribonucleoside kinase with a K(m) of 4.5 microM and a V(max) of 400 nmol/microg protein/h compared with 1.3 microM and 62.5 nmol/microg protein/h, respectively, for the natural substrate thymidine. Mammalian cytosolic thymidine kinase-1 does not recognize BVDU as a substrate. In sharp contrast to mammalian cells, the insect D. melanogaster and Spodoptera frugiperda (Sf) embryonic cells proved highly sensitive to the cytostatic action of BVDU. BVDU was efficiently metabolized to its 5'-mono-, 5'-di- and 5'-triphosphate derivatives in the insect cell cultures and abundantly incorporated into the insect cell DNA. BVDU prevented the D. melanogaster cells to initiate the S phase of their cell cycle, and exposure of S. frugiperda cells to BVDU led to a dose-dependent retardation of the insect cells in the S phase of their cell cycle. Both inhibition of nucleic acid synthesis (through the 5'-triphosphate of BVDU) and inhibition of thymidylate synthase (through the 5'-monophosphate of BVDU) would account for the cytostatic activity of BVDU against the insect cells. Because of the virtual lack of cytotoxicity of BVDU against mammalian cells, the drug should be considered highly selective in its cytostatic action against the insect cells. When added to the food of S. frugiperda larvae, BVDU caused a remarkable decrease in the weight gain of the larvae and heavily compromised the transformation of the larvae to the pupae and their subsequent adult (moth) phase. Our data indicate that insect multifunctional deoxyribonucleoside kinase should be considered an entirely novel and attractive target in the development of new nucleoside types of highly selective insecticidal drugs.

Animals↗
PubMed1999

Respiratory burst in human neutrophils.

During phagocytosis of microbial intruders, professional phagocytes of our innate immune system increase their oxygen consumption through the activity of an NADPH-oxidase that generates superoxide anion (O(2)(-)) and hydrogen peroxide (H(2)O(2)). These oxygen metabolites give rise to yet other reactive oxygen species that are strongly anti-microbial but which may also cause damage by destructing surrounding tissue and inducing apoptosis in other immune reactive cells. The development of methodology to measure the generation/release of phagocyte respiratory burst products is thus of great importance, and a number of different techniques are currently in use for this purpose. Three of the techniques that we have used, (luminol/isoluminol amplified chemiluminescence, cytochrome C reduction, and PHPA oxidation technique) are described in more detail in this review. We hope to convince the readers that these techniques are valuable tools in basic as well as more clinically oriented research dealing with phagocyte function. The basic principles for luminol/isoluminol-amplified chemiluminescence is used as the starting point for discussing methodological problems related to measurements of oxygen metabolites generated by professional phagocytes.

Fluorometry↗
PubMed1999

Identification of CD66a and CD66b as the major galectin-3 receptor candidates in human neutrophils.

The mammalian lectin galectin-3 is a potent stimulus of human neutrophils, provided that the receptor(s) for the lectin has been mobilized to the cell surface before activation. We have recently shown that the receptors for galectin-3 are stored in intracellular mobilizable granules. Here we show supportive evidence for this in that DMSO-differentiated (neutrophil-like) HL-60 cells, which lack gelatinase and specific granules, are nonresponsive when exposed to galectin-3. Neutrophil granules were subsequently used for isolation of galectin-3 receptors by affinity chromatography. Proteins eluted from a galectin-3-Sepharose column by lactose were analyzed on SDS-polyacrylamide gels and showed two major bands of 100 and 160 kDa and a minor band of 120 kDa. By immunoblotting, these proteins were shown to correspond to CD66a (160 kDa), CD66b (100 kDa), and lysosome-associated membrane glycoprotein-1 and -2 (Lamp-1 and -2; 120 kDa). The unresponsive HL-60 cells lacked the CD66 Ags but contained the Lamps, implying that neutrophil CD66a and/or CD66b may be the functional galectin-3 receptors. This conclusion was supported by the subcellular localization of the CD66 proteins to the gelatinase and specific granules in resting neutrophils.

Antigens, CD↗
PubMed1999

Packed-column supercritical fluid chromatography for the purity analysis of clevidipine, a new dihydropyridine drug.

In this paper we describe a packed column supercritical fluid chromatography method that can be used for the analysis of a new dihydropyridine substance. The method is based on methanol-modified carbon dioxide as the mobile phase and Hypersil bare silica as column support at a column temperature of 50 degrees C and 150 bar as back pressure. Using an adjusted methanol gradient the most likely by-products can be separated and detected (240 nm) within 13 min. Occasionally the column needed treatment with 4 mM citric acid in the methanol modifier in order to give a narrow peak of an acidic analogue. The present method can detect analogues at the 0.1% (w/w) level. The precision at this level for one of the analogues was 5.9% RSD. This method shows a higher selectivity than a corresponding reversed-phase liquid chromatographic method.

Antihypertensive Agents↗
PubMed1999

Cloning of mouse mitochondrial thymidine kinase 2 cDNA.

Phosphorylation of anti-viral nucleoside analogs by mitochondrial thymidine kinase 2 (TK2) has been implicated as a mechanism for the mitochondrial toxicity caused by several of these compounds. We have cloned the cDNA of mouse TK2 and shown that the enzyme is targeted to the mitochondria when expressed in fusion with the green fluorescent protein. This is the first report on the cloning of a mitochondrial TK2 and will contribute to elucidate the role of TK2 in the pharmacological activation of nucleoside analogs.

Amino Acid Sequence↗
PubMed1999

5'-Phosphoramidates and 5'-diphosphates of 2'-O-allyl-beta-D-arabinofuranosyluracil, -cytosine, and -adenine: inhibition of ribonucleotide reductase.

Continuing our studies on ribonucleotide reductase (RNR) mechanism-based inhibitors, we have now prepared the diphosphates (DP) of 2'-O-allyl-1-beta-D-arabinofuranosyl-uracil and -cytosine and 2'-O-allyl-9-beta-D-arabinofuranosyl-adenine and evaluated their inhibitory activity against recombinant murine RNR. 2'-O-Allyl-araUDP proved to be inhibitory to RNR at an IC(50) of 100 microM, whereas 2'-O-allyl-araCDP was only marginally active (IC(50) 1 mM) and 2'-O-allyl-araADP was completely inactive. The susceptibility of the parent nucleosides to phosphorylation by thymidine kinase and 2'-deoxycytidine kinase was also investigated, and all nucleosides proved to be poor substrates for the above-cited kinases. Moreover, prodrugs of 2'-O-allyl-araU and -araC monophosphates, namely 2'-O-allyl-5'-(phenylethoxy-L-alanyl phosphate)-araU and -araC, were prepared and tested against tumor cell proliferation but proved to be inactive. A molecular modeling study has been conducted in order to explain our results. The data confirm that for both the natural and analogue nucleoside diphosphates, the principal determinant interaction with the active site of RNR is with the diphosphate group, which forms strong hydrogen bonds with Glu623, Thr624, Ser625, and Thr209. Our findings indicate that the poor phosphorylation may represent an explanation for the lack of marked in vitro cytostatic activity of the test compounds.

Animals↗
PubMed1999

Cloning and characterization of the multisubstrate deoxyribonucleoside kinase of Drosophila melanogaster.

A Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) was reported to phosphorylate all four natural deoxyribonucleosides as well as several nucleoside analogs (Munch-Petersen, B., Piskur, J., and Sondergaard, L. (1998) J. Biol. Chem. 273, 3926-3931). The broad substrate specificity of this enzyme together with a high catalytic rate makes it unique among the nucleoside kinases. We have in the present study cloned the Dm-dNK cDNA, expressed the 29-kDa protein in Escherichia coli, and characterized the recombinant enzyme for the phosphorylation of nucleosides and clinically important nucleoside analogs. The recombinant enzyme preferentially phosphorylated the pyrimidine nucleosides dThd, dCyd, and dUrd, but phosphorylation of the purine nucleosides dAdo and dGuo was also efficiently catalyzed. Dm-dNK is closely related to human and herpes simplex virus deoxyribonucleoside kinases. The highest level of sequence similarity was noted with human mitochondrial thymidine kinase 2, and these enzymes also share many substrates. The cDNA cloning and characterization of Dm-dNK will be the basis for studies on the use of this multisubstrate nucleoside kinase as a suicide gene in combined gene/chemotherapy of cancer.

Amino Acid Sequence↗
PubMed1999

Chemical transformations in individual ultrasmall biomimetic containers.

Individual phospholipid vesicles, 1 to 5 micrometers in diameter, containing a single reagent or a complete reaction system, were immobilized with an infrared laser optical trap or by adhesion to modified borosilicate glass surfaces. Chemical transformations were initiated either by electroporation or by electrofusion, in each case through application of a short (10-microsecond), intense (20 to 50 kilovolts per centimeter) electric pulse delivered across ultramicroelectrodes. Product formation was monitored by far-field laser fluorescence microscopy. The ultrasmall characteristic of this reaction volume led to rapid diffusional mixing that permits the study of fast chemical kinetics. This technique is also well suited for the study of reaction dynamics of biological molecules within lipid-enclosed nanoenvironments that mimic cell membranes.

Alkaline Phosphatase↗
PubMed1999

Degradation of phenol under meso- and thermophilic, anaerobic conditions.

Based on the results of preliminary studies on phenol degradation under mesophilic conditions with a mixed methanogenic culture, we proposed a degradation pathway in which phenol is fermented to acetate: Part of the phenol is reductively transformed to benzoate while the rest is oxidised, forming acetate as end product. According to our calculations, this should result in three moles of phenol being converted to two moles of benzoate and three moles of acetate (3 phenol + 2 CO2 + 3 H2O --> 3 acetate + 2 benzoate): To assess the validity of our hypothesis concerning the metabolic pathway, we studied the transformation of phenol under mesophilic and thermophilic conditions in relation to the availability of hydrogen. Hence, methanogenic meso- and thermophilic cultures amended with phenol were run with or without an added over-pressure of hydrogen under methanogenic and non-methanogenic conditions. Bromoethanesulfonic acid (BES) was used to inhibit methanogenic activity. In the mesophilic treatments amended with only BES, about 70% of the carbon in the products found was benzoate. During the course of phenol transformation in these BES-amended cultures, the formation pattern of the degradation products changed: Initially nearly 90% of the carbon from phenol degradation was recovered as benzoate, whereas later in the incubation, in addition to benzoate formation, the aromatic nucleus degraded completely to acetate. Thus, the initial reduction of phenol to benzoate resulted in a lowering of H2 levels, giving rise to conditions allowing the degradation of phenol to acetate as the end product. Product formation in bottles amended with BES and phenol occurred in accordance with the hypothesised pathway; however, the overall results indicate that the degradation of phenol in this system is more complex. During phenol transformation under thermophilic conditions, no benzoate was observed and no phenol was transformed in the BES-amended cultures. This suggests that the sensitivity of phenol transformation to an elevated partial pressure of H2 is higher under thermophilic conditions than under mesophilic ones. The lack of benzoate formation could have been due to a high turnover of benzoate or to a difference in the phenol degradation pathway between the thermophilic and mesophilic cultures.

Journal Article↗
PubMed1999

A new bone dust packer for use in neurosurgery.

A new tool for the packing of bone dust has been developed. The instrument compresses a mixture of bone dust and saline into suitable cylindrical plugs, fitting neatly into craniotomy burrholes. The skullbone at the craniotomy area will heal neatly also assuring the best healing possibilities for the overlying skin. The cosmetic results are excellent as no skin dents or cavities will be formed over the burrholes.

Adult↗
PubMed1999

In vivo cell recruitment, cytokine release and chemiluminescence response at gold, and thiol functionalized surfaces.

Hydroxylated and methylated surfaces were prepared by the self-assembled monolayer technique (SAM) of alkane thiols on gold. The surfaces were used to evaluate the influence of implant surface chemistry on protein deposition and inflammatory cell response. Implants were inserted subcutaneously in the rat for 3 and 24 h. The surface chemical properties influenced the in vitro rat plasma protein adsorption (ellipsometry/antibody) with few exceptions (albumin not found and fibrinogen always found). The number of recruited cells and their distribution (DNA from implant versus from exudate) was influenced by the different chemistries at 24 h, but not at 3 h. HIS48+, ED1+, ED2+ and small numbers of CD5+ cells were present in the exudate at both time periods (flow cytometry). The cellular oxidative metabolism was low, although cells on -OH surfaces responded with the highest phorbol ester-stimulated chemiluminescence (CL)/DNA. The levels of cytokines IL-1alpha, IL-1beta and TNFalpha (ELISA) were not influenced by material surface chemistry. Sham operated sites had a higher cytokine concentration/DNA compared with exudates from an implant milieu. The results of this study show that surface chemical functionalization modifies specific events in the inflammatory response around implants in soft tissues.

Adsorption↗
PubMed1999

Enantiometric separation of verapamil and norverapamil using Chiral-AGP as the stationary phase.

Simultaneous enantiomeric separation of verapamil and its main metabolite norverapamil was achieved using Chiral-AGP as the stationary phase. The optimized chromatographic system was obtained using statistical experimental design with partial least squares as regression method. The three variables studied were buffer pH, content of acetonitrile and column temperature. A high buffer pH favors enantioselectivity as well as the selectivity between (S)-verapamil and (R)-norverapamil. The concentration of the organic modifier in the mobile phase was a compromise as a high content of acetonitrile decreased enantioselectivity but increased the selectivity mentioned above. Increased column temperature increased the separation between (S)-verapamil and (R)-norverapamil with only a slight decrease in enantioresolution.

Calcium Channel Blockers↗