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Biomedical subjects

K Mortensen

Publications and source records attributed to K Mortensen.

At least 19 recordsLinked to original sources

Collective dynamics and self-diffusion in a diblock copolymer melt in the body-centered cubic phase.

The structure and dynamics of a strongly asymmetric poly(ethylene propylene)-poly(dimethylsiloxane) (PEP-PDMS) diblock copolymer in the melt have been studied over a wide temperature range. Small-angle neutron scattering reveals that the sample exhibits two stable phases in this temperature range: Above the order-to-disorder transition temperature, it is disordered, whereas the domain structure is body-centered cubic (bcc) below, being stable down to the lowest temperatures measured. In the disordered state, dynamic light scattering (DLS) in the polarized geometry reveals the heterogeneity mode and the cluster mode. In the bcc phase, the PEP and the PDMS blocks form the micellar cores and the matrix, respectively. Here, two modes are observed in DLS, and the diffusion coefficients measured using pulsed field gradient (PFG) NMR are broadly distributed with the most probable diffusion coefficient coinciding with the slow DLS mode. We attribute the fast process in the bcc state to concentration fluctuations of the micellar cores (PEP), relaxing by mutual diffusion of the micelles with copolymers dissolved in the PDMS matrix. The slower process in the bcc state is ascribed to activated long-range self-diffusion of single copolymers from micelle to micelle through the PDMS matrix. This assignment is corroborated by the good coincidence of the reduced diffusivities with the ones from the literature. However, this mode may also be assigned to the rearrangement of entire micelles.

Journal Article↗

Spontaneous fusion between cancer cells and endothelial cells.

Endothelial cells line the inside of blood and lymphatic vessels, and cancer cells must cross this barrier, first to gain access to the circulation, and, second, to exit and metastasize. How this occurs is incompletely understood. We now demonstrate that human cancer cells are able to fuse with endothelial cells to form hybrid cells displaying proteins and chromosomal markers characteristic of both parent cells. The hybrid cells are viable and capable of undergoing mitosis. Fusions between cancer cells and endothelial cells were shown to occur both in vitro, in co-cultures of human breast cancer cells and endothelial cells, and in vivo, following intravascular dissemination of human breast cancer cells in nude mice. These observations demonstrate a new type of cancer-endothelial cell interaction that may be of fundamental importance to the process of metastasis.

Animals↗

Effects of cytochalasin D on the actin cytoskeleton: association of neoformed actin aggregates with proteins involved in signaling and endocytosis.

Cytochalasin D (CD) has been extensively used for assessing the role of the actin cytoskeleton in different biological processes. However, effects of CD have not always been consistent and CD-treated cells have been found to contain irregular spots of F-actin. By transfecting MCF-7 cells with an actin-enhanced yellow fluorescent protein fusion protein we show that, in vivo, CD induces actin aggregation de novo, while simultaneously depolymerizing preexisting actin cytoskeletal components. We also show that CD-induced actin aggregates bind the F-actin-selective drug phalloidin and associate with proteins involved in cell signaling as well as with receptors and endosomal markers (active MAP kinases, paxillin, erbB2, transferrin, Rab-5), but not with clathrin, protein kinase A, protein tyrosine phosphatase 1B, or tubulin. Thus, CD induces new sites of actin aggregation that selectively associate with several important regulatory proteins. Failure of CD to interupt a biological process may therefore not prove that the process is independent of actin aggregation.

Actins↗

Neuronal nitric oxide synthase activity is increased during granulomatous inflammation in the colon and caecum of pigs infected with Schistosoma japonicum.

Neuronal nitric oxide is a non-adrenergic non-cholinergic neurotransmitter in the enteric nervous system and plays a role in a variety of enteropathies including Crohn's and Chagas' diseases, ulcerative colitis, diabetes, atrophy and hypertrophy. The content of neuronal nitric oxide synthase (nNOS) in the colon and the caecum from pigs infected with Schistosoma japonicum was studied using immunohistochemical and histochemical staining for nNOS and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-diaphorase), respectively. In the infected pigs, lightly, moderately and less severely inflamed tissues showed increased nNOS and NADPH-diaphorase activities in nerve cell bodies and nerve fibres in the enteric plexuses compared to control pigs. There was a significant increase in the nerve cell body density of nNOS immunoreactive nerve cell bodies in the inner submucous plexus, outer submucous plexus and in the myenteric plexus. More intensely stained nerve cell bodies and varicosities were observed in tissue from prenatally infected and prenatally infected, postnatally re-infected pigs compared to postnatally infected pigs. However, the latter showed the highest numerical density of nNOS immunoreactive nerve cell bodies. Marked increases were seen in the inner submucous plexus followed by myenteric plexus, inner circular muscle, outer submucous plexus and mucous plexus. However, in very severe inflamed tissues, the number and staining intensity of nerve cell bodies and nerve fibre varicosities were reduced in plexuses located in the lesions with the inner submucous and mucous plexuses being the most affected. There was no staining in the nervous tissue within the eosinophilic cell abscesses and productive granulomas. The apparent alterations in the activities of enzymes responsible for the generation of nitric oxide (NO) show possible alterations in the NO mediated non-adrenergic non-cholinergic reflexes in the enteric nervous tissue. These alterations might contribute to impaired intestinal motility and absorption, and other pathophysiological conditions seen during S. japonicum infections.

Animals↗

Temperature dependence of the flux line lattice transition into square symmetry in superconducting LuNi2B2C.

We have investigated the temperature dependence of the H parallel to c flux line lattice structural phase transition from square to hexagonal symmetry, in the tetragonal superconductor LuNi2B2C ( T(c) = 16.6 K). At temperatures below 10 K the transition onset field, H2(T), is only weakly temperature dependent. Above 10 K, H2(T) rises sharply, bending away from the upper critical field. This contradicts theoretical predictions of H2(T) merging with the upper critical field and suggests that just below the H(c2)(T) curve the flux line lattice might be hexagonal.

Journal Article↗

Flux line lattice reorientation in the borocarbide superconductors with H parallel a.

Small angle neutron scattering studies of the flux line lattice in LuNi2B2C and ErNi2B2C induced by a field parallel to the a axis reveal a first order flux line lattice reorientation transition. Below the transition the flux line lattice nearest neighbor direction is parallel to the b axis, and above the transition it is parallel to the c axis. This transition cannot be explained using nonlocal corrections to the London model. In addition, the anisotropy of the penetration depth lambda and the coherence length xi change at the transition.

Journal Article↗

Quantitative and qualitative immunofluorescence studies of neoplastic cells transfected with a construct encoding p53-EGFP.

The p53 protein is a major regulator of cell cycle progression and apoptosis. We used a p53-enhanced green fluorescent protein (EGFP) construct for transfections into human breast cancer (MCF-7) cells. Cells expressing p53-EGFP showed an increased apoptotic index compared to cells transfected with EGFP alone. Interestingly, apoptotic cells showed localization of p53-EGFP to both nuclei and cytoplasm, whereas non-apoptotic cells usually only showed nuclear localization of p53-EGFP. This result is in agreement with the hypothesis that p53 induces apoptosis by interaction with both nuclear and cytoplasmic targets. Transfected p53-deficient osteosarcoma cells were used for immunofluorescence quantitation. The intensity of immunofluorescence for either p53 or EGFP showed excellent linear correlation to the EGFP autofluorescence, proving that measurements of immunofluorescence intensities can be used for determining endogenous protein levels.

Fluorescent Antibody Technique, Indirect↗

Reversible thermal gelation in soft spheres.

Upon heating, concentrated solutions of star polymers and block copolymer micelles in a good solvent, representing soft spheres, undergo a reversible gelation. This phenomenon is attributed to the formation of clusters causing a partial dynamic arrest of the swollen interpenetrating spheres at high temperatures. A phase diagram analogous to that of sterically stabilized colloids is proposed.

Journal Article↗

Apoptosis in rat gastric antrum: evidence that regulation by food intake depends on nitric oxide synthase.

The turnover of the epithelium of the gastrointestinal tract is regulated by a balance between cell multiplication and cell loss. We examined the effects of starvation on apoptosis in endocrine and other epithelial cells of rat antropyloric mucosa. Apoptosis was determined by the TUNEL reaction combined with immunocytochemical staining for gastrin and somatostatin. Apoptotic cell morphology was determined by bisbenzimide staining for DNA. Both gastrin and somatostatin cells showed a significantly lower apoptotic index than the general epithelium. This agrees with the longer turnover kinetics of gastric endocrine cells. On starvation, the apoptotic index of the general epithelium and of the gastrin but not of the somatostatin, cells increased significantly. This was prevented by the nitric oxide synthase (NOS) inhibitor L-NAME but not by its inactive stereoisomer D-NAME. Immunoreactive neuronal NOS was present in somatostatin cells, in nonendocrine cells predominating in the surface and pit epithelium, and in rare nerve fibers. Endothelial cell NOS was present in vessels, whereas the inducible isoform was barely detectable. Thus, endogenous NOS isoforms participate in regulating antropyloric epithelial apoptosis during starvation. The close paracrine relation between somatostatin cells and gastrin cells suggests that the former regulates apoptosis of the latter through release of NO.

Animals↗

Endogenous endothelial cell nitric-oxide synthase modulates apoptosis in cultured breast cancer cells and is transcriptionally regulated by p53.

Nitric oxide can both stimulate and suppress apoptosis. By reverse transcriptase-polymerase chain reaction and sequencing we show that human breast cancer (MCF-7) cells express endothelial cell nitric-oxide synthase (ecNOS), but not other nitric-oxide synthase isoforms. Inhibition of ecNOS activity in MCF-7 cells increased tumor cell apoptosis, and this effect was also seen following treatment with an NO scavenger. In addition, low concentrations of the NO donor sodium nitroprusside inhibited, whereas high concentrations stimulated MCF-7 cell apoptosis. The ecNOS promoter was found to contain a specific binding site for the apoptosis-regulating protein p53. In co-transfection studies wild-type, but not mutant, p53 down-regulated transcription of an ecNOS promoter-luciferase reporter gene construct. In addition, NO donors up-regulated p53 protein levels in MCF-7 cells. These data point to a previously unrecognized p53-dependent regulation of ecNOS expression that may be important both for regulating apoptosis and for avoiding the generation of genotoxic quantities of NO.

Apoptosis↗

[Epidemiology of head injuries in Troms].

This retrospective population-based survey describes the epidemiology of head injury in a defined population in Troms. It includes all 247 patients with head injury referred to the University Hospital of Tromsø during 1993. Head injury was defined as physical damage to the brain or skull caused by external force. The annual incidence rate of hospital-referred head injury was 229/100,000 population with a male preponderance of 1.7:1.0. Causes were fall in 62%, traffic accident in 21%, and assault in 7% of the cases. The observed incidence rate is low despite the use of wide inclusion criteria, probably due to a decrease in road traffic accidents and fewer referrals. A further decrease in the number of head injuries in our region may be achieved by preventing falls.

Adolescent↗

[Management of minor head injuries in Norwegian hospitals--can the quality be improved?].

Management protocols for minor head injury should include strategies for early detection of intracranial haematomas. This study focuses on the management of minor head injury in 63 Norwegian hospitals. We report considerable inter-hospital variation. In most (81%) hospitals, minor head injury patients were treated by general surgeons. Emergency room evaluation included routine radiological evaluation, usually skull radiography, in 18 (29%) hospitals, and assessment according to the Glasgow Coma Scale (GCS) in 27 (43%). GCS was used during in-hospital observation in 32 (51%) hospitals. 33 (52%) discharged selected minor head injury patients without in-hospital observation. We conclude that the quality of care for minor head injury patients in Norwegian hospitals can be improved through extended use of routine early CT and consistent evaluation according to GCS.

Brain Injuries↗

Difference between active and inactive nucleotide cofactors in the effect on the DNA binding and the helical structure of RecA filament dissociation of RecA--DNA complex by inactive nucleotides.

The RecA protein requires ATP or dATP for its coprotease and strand exchange activities. Other natural nucleotides, such as ADP, CTP, GTP, UTP and TTP, have little or no activation effect on RecA for these activities. We have investigated the activation mechanism, and the selectivity for ATP, by studying the effect of various nucleotides on the DNA binding and the helical structure of the RecA filament. The interaction with DNA was investigated via fluorescence measurements with a fluorescent DNA analog and fluorescein-labeled oligonucleotides, assisted by linear dichroism. Filament structure was investigated via small-angle neutron scattering. There is no simple correlation between filament elongation, DNA binding affinity of RecA, and DNA structure in the RecA complex. There may be multiple conformations of RecA. Both coprotease and strand exchange activities require formation of a rigid and well organized complex. The triphosphate nucleotides which do not activate RecA, destabilize the RecA-DNA complex, indicating that the chemical nature of the nucleotide nucleobase is very important for the stability of RecA-DNA complex. Higher stability of the RecA-DNA complex in the presence of adenosine 5'-O-3-thiotriphosphate or guanosine 5'-O-3-thiotriphosphate than ATP or GTP indicates that contact between the protein and the chemical group at the gamma position of the nucleotide also affects the stability of the RecA-DNA complex. This contact appears also important for the rigid organization of DNA because ADP strongly decreases the rigidity of the complex.

Circular Dichroism↗

Endothelial cell nitric oxide synthase in peritumoral microvessels is a favorable prognostic indicator in premenopausal breast cancer patients.

Nitric oxide (NO) is involved in tumor cell apoptosis and has additional effects on tumor blood flow, immune responses, and angiogenesis. We, therefore, studied endothelial cell NO synthase (ecNOS) protein expression in a retrospective series of 118 patients with primary invasive breast cancer. Immunocytochemically stained paraffin sections were used for determining the frequency of (a) tumor cells, (b) intratumoral microvessels, and (c) peritumoral microvessels that were positive for ecNOS. A high density of ecNOS positive microvessels in the normal tissue surrounding the tumor (measured by the variable PEMVD) was associated with significantly better recurrence-free and overall survival. The prognostic significance was observed in a representative series of premenopausal patients and was independent of other factors, including lymph node status. The counting procedure was highly reproducible and correlated to stereological measurements but was influenced by heterogeneity of the tissue samples. Analyzing two sections per patient improved the discriminative power by reducing the influence of tissue heterogeneity and produced highly significant results (recurrence-free survival, P < 0.001; overall survival, P < 0.0001). Immunoreactive ecNOS in microvessels is an independent prognostic factor in breast cancer and may reflect a mechanism of endothelial defense against invasion by tumor cells. Individual variations in ecNOS may be related to environmental, hormonal, and genetic factors and could represent a therapeutic target.

Adult↗

Interfacial Modification as a Route to Novel Bilayered Morphologies in Binary Block Copolymer/Homopolymer Blends.

Addition of a relatively low-molecular-weight parent homopolymer to a lamellar AB diblock copolymer constitutes a reliable means by which to induce, in controllable fashion, transitions to other morphologies. In this study, we examine the effect of interfacial modification on such transitions in "extended" A(A/B)B copolymer/homopolymer blends in which (i) the A/B midblock fraction (relative to the copolymer molecular weight) is varied from 0.0 to 0.4 in 0.1 increments and (ii) the overall concentration of A ranges from 0.50 to 0.95. As this A/B fraction is increased at constant blend composition, the extent of homopolymer-induced lamellar swelling becomes measurably less pronounced, indicating that the A/B midblock serves to delocalize repulsion along the interphase separating adjacent lamellae. At higher homopolymer concentrations, an increase in the A/B fraction results in the formation of either unilamellar vesicles or a randomly connected bilayered membrane, rather than micelles. These membranes become unstable and transform to micelles at high copolymer dilution. The results presented here are discussed in terms of the complex morphologies observed in, and predicted for, low-molar-mass (co)surfactant systems.

Journal Article↗

Influence of Alcohol on the Behavior of Sodium Dodecylsulfate Micelles.

The effect of medium chain alcohol molecules on the size and shape of sodium dodecylsulfate micelles, and on the self-diffusion coefficient of the surfactant and alcohol, has been investigated by means of small angle neutron scattering (SANS), and Fourier transform pulsed field gradient spin echo (FT-PGSE) nuclear magnetic resonance measurements. All measurements were done in D2O containing a sodium chloride concentration of 0.4 mol/kg, and a surfactant concentration of 0.04 mol/kg. The alcohols used were 1-propanol, 1-butanol, and 1-pentanol. The data obtained from the different techniques agrees qualitatively. The results show that propanol successively breaks down the micelles while pentanol brings about a structural change toward large wormlike aggregates. Butanol shows a highly complex behavior on the structure of the micelles and can decrease and increase the size of the aggregates, depending on the added alcohol concentration range. All analyzed solutions show a distribution of the alcohol between the aqueous bulk solution and the palisade layer of the micelles, resulting in an increased "hydration" of D2O in the micellar surface. Moreover, the structural changes of the micelles indicate that butanol and pentanol solubilize in the micellar core in the high molality range, near the end of the solubility limit. Propanol is apparently too hydrophilic to do so. Copyright 1998 Academic Press.

Journal Article↗

Base orientation of second DNA in RecA.DNA filaments. Analysis by combination of linear dichroism and small angle neutron scattering in flow-oriented solution.

To gain insight into the mechanism of pairing two complementary DNA strands by the RecA protein, we have determined the nucleobase orientation of the first and the second bound DNA strands in the RecA.DNA filament by combined measurements of linear dichroism and small angle neutron scattering on flow-oriented samples. An etheno-modified DNA, poly(depsilonA) was adapted as the first DNA and an oligo(dT) as the second DNA, making it possible to distinguish between the linear dichroism signals of the two DNA strands. The results indicate that binding of the second DNA does not alter the nucleobase orientation of the first bound strand and that the bases of the second DNA are almost coplanar to the bases of the first strand although somewhat more tilted (60 degrees relative to the fiber axis compared with 70 degrees for the first DNA strand). Similar results were obtained for the RecA.DNA complex formed with unmodified poly(dA) and oligo(dT). An almost coplanar orientation of nucleobases of two DNA strands in a RecA-DNA filament would facilitate scanning for, and recognition of, complementary base sequences. The slight deviation from co-planarity could increase the free energy of the duplex to facilitate dissociation in case of mismatching base sequences.

DNA↗